Innovative Solutions for Flow Cytometry Analysis Optimized Kits and Reagents PB3322ENEU

8/3/2019 Innovative Solutions for Flow Cytometry Analysis Optimized Kits and Reagents PB3322ENEU

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Innovative Solutions orFlow Cytometry Analysis

Optimized Kits and Reagents


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Flow cytometry is an essential tool or in-depth cell

analysis. With the capacity to simultaneously

measure multiple parameters on hundreds o

individual cells per second, ow cytometry oers

greater speed, precision, and detail than most other

cell analysis methods available to scientists today.

Integrated products rom Merck Millipore will help

streamline your workow. Our complete benchtop

ow cytometry solution includes our instruments,

assays, and sotwareas well as the cell

isolation and culture tools to prepare your samples.

With a ow cytometry solution right in your lab,

youll experience superior perormance, higher

quality data and aster progress rom hypothesis to

results.

Guava integratedow cytometry solutions

Whats inside...

Cell Health Cell Counting & Viability

Cell Cycle

DNA Damage

Mitochondrial Analysis

Apoptosis

Cell Signaling MAPK Pathway

EGFR Pathway

PI3/Akt/mTOR Pathway

Jak/STAT Pathway

Chemokine

Stem Cells Embryonic Stem Cell

(Human/Mouse)

Neural Stem Cell

(Rodent)

Immunology Regulation T-Cell

Phenotype Markers

Milli-MarkConjugatedAntibodies

Componentso the guavaFlow CytometrySolution Instruments: easyCyte

Flow Cytometers

Sotware

Kits & Reagents


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Kits and Reagents

FlowCellect Kits and

Milli-Mark Conjugated

Antibodies FlowCellect Kits

Milli-Mark Conjugated

Antibodies

Many researchers invest time

optimizing and validating antibodies

or ow cytometry, only to discover

that these antibodies do not perorm

well when multiplexed together,

because o intererence rom the

matrix or rom other antibodies.

Merck Millipores FlowCellect kits

and Milli-Mark conjugated primary

antibodies are ully optimized

or ast, easy, and accurate

multiparametric ow cytometry.

Weve taken the guesswork out o

assay development so you can ocus

on your results. We optimize and

validate every antibody, making surethey work together well. All you need

are cells and a research question; our

assay kits will do the rest, and youll

have data beore your cells are ready

to split again.

FlowCellect kits are Merck Millipores

proprietary multiparameter ow

cytometry kits or the analysis

o cellular events and/or cell

phenotypes. Each kit has unique

combination o directly conjugated

antibodies, and/or uorescent dyes

and protein reporters to monitor

changes in protein expression and

posttranslational modifcation. The

kits also contain complete buer

sets, protocols and pre-defned gate

settings. They are ully optimized or

plug-and-play cellular analysis on

guava instruments and other ow

cytometers.

Using a our-step validation process

to develop our FlowCellect kits, weve

eliminated the need to design your

experiment or optimize antibodies

and buers (Figure 2 below).

Milli-Mark antibodies are directly

conjugated primary antibodies

that are validated or ow

cytometry in addition to traditional

applications like Western blotting

and immunocytochemistry (see

Figure 1B or an example o antibody

validation). Because o their

extensive crossplatorm validation,

Milli-Mark antibodies are valuable,

convenient building blocks with

which you can confgure your own

assays.

The antibodies used in

each kit are careully

selected by reviewing

many publications.

Antibodies are conjugated

to compatible

uorophores that will

ensure less uorescence

spectrum overlap.

Antibodies are screened

primarily by Western blot

to determine specifcity,

then validated or ow

cytometry using a

secondary antibody.

The antibody conjugates

are optimized to provide

the best signal-to-noise

ratio when multiplexing.

Figure 2. FlowCellect Kit Four-step Validation Process.

Antibody Selection

and Assay Design

Antibody Conjgation

and Testing

Antibody Component

Validation

Mltiplexing,

Stability, Perormance

Claims

Figure1. (A) Mouse embryonicstem cells and fbroblastslabeled with Oct-4 (green),SSEA-1 (red), and Hoechstnuclear stain (blue). (B) Humanembryonic stem cell lysate.

Advantages

Multiplexingcapabilities

Easytouse,withfewerincubationandwashsteps

Fullyvalidatedandconcentration-optimized,guaranteed

to work in ow cytometry

B.

A.

OCT-4antibody

SSEA-4antibody

250

130

100

70

55

35

27

15

10

250

130

100

70

55

35

27

15

10

44kDa

70kDa


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Cell Health

Knowing the perormance profle o your cells prior to

running your bioassay can mean the di erence between

valid assay results and wasted reagents, lost time and

discarded data.

Monitoring key indicators o cell health and perormance,

such as the apoptotic raction and stage o apoptosis,

viability, cell cycle, cell counts, transection efciency, and

target expression levels, helps establish uniorm standards

o cellular perormance across long-term research studies.

These standards can be applied to a wide range obioassays. Whether you are establishing screen/no screen

criteria or high throughput screening, monitoring and

optimizing bioreactor conditions, or eliminating sources o

assay variability, consistent monitoring o your cell model

improves your bioassay perormance and productivity.

Cell Conting and ViabilityCell counting and viability assessments can be used in a

variety o applications, such as cytotoxicity studies, PBMC

counting and rapid apoptosis assessment.

gava ViaCont Assay Kits

The guava ViaCount assay is ast becoming the new

standard or viability and cell counting. In this simple

no-wash, mix-and-read-assay, you can accurately obtain

absolute total cell counts, perorm viability assessments

and determine apoptotic percentagesall rom tiny

samples. Youll enjoy several advantages over traditional

methods, including greater accuracy, reproducibility and

speed.

Advantages

Assays:

Simpleno-wash,mix-and-readprocedure

Countsupto10timesfasterthanmanualmethods

Morereproduciblethantraditionaltests

Samples:

Usessmallsamplesintubesora96-wellplate

Handleslow-densityandsmall-volumecellsamples Workswithadherentorsuspendedcellsandmammalian

and insect cells

Description Qty Catalog No.

Guava ViaCount Reagent Kit

The new standard or cell counting and viability assessment.

100tests 4000-0040

600tests 4000-0041

Guava ViaCount Flex Reagent Kit

Non-mammaliancelllines(ie.SF9insectcells).

Lowdensitycellsamples(~104 cells/mL).

Cell lines that stain heterogeneously.

100tests 4500-0110

500tests 4700-0060

Guava ViaCount Cell Dispersal Reagent Kit (CDR)

Uses enzymes to gently disaggregate clumped cells in suspension, improve the accuracy

and precision o cell counts.

100tests 4700-0050

The blue population o cells show a signifcant amount o

annexin V staining (right plot), indicating that intermediate

levels o staining with the viability dye correlates with apoptosis.

Discover the power o ow cytometry

or multiparametric cell health analysis

Viability Stain

Nucle

ated

Cells

Live

Apoptotic

Dead

Annexin V

Live

Apoptotic

Dead

104

103

102

101

100100 101 102 103 104

104

103

102

101

100100 101 102 103 104

ViabilityStain

gava ViaCont Assay Kits


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Cell CycleThe cell cycle can be divided into two distinct stages. The

frst stage is interphase which consists o the G1, S, and

G2 phases, in which cells are active, growing, and DNA is

being replicated. The second is M phase, also known as the

mitotic phase, in which cell division takes place.

Cell cycle phase distributions can be used to assess cell

health, prolieration, as well as the potential mechanism

o antineoplastic agents. For example, measuring the

population o S phase cells can reect the amount o newly

synthesized DNA. Also, distinguishing cells in G2 rom M

phase cells can help identiy cells undergoing mitosis.

Flow cytometry analysis o cell DNA content has been

one o the best and most popular tools or researchers

to be widely used or the estimation o cell cycle phase

distribution. However, the limitation o single-marker

analysis, such as a DNA dye only, is that cells within

each phase cannot accurately be determined without

mathematical interpolation using analysis sotware.

FlowCellect Cell Cycle Kits

Merck Millipore has developed and optimized two bivariate

cell cycle analysis kits using phase-specifc antibodies

in addition to a DNA dye. Bivariate analysis will not only

reveal the cell distribution within a particular phase o

cell cycle, but can also enable the researcher to elucidate

mechanisms o cell cycle regulation, without sophisticated

sotware modules or algorithms.

Advantages

Quantitative

measurements opercentage o cells withineach cell subpopulation

Minimalassaydevelopment needed

Includesalloptimizedow cytometry antibodiesand buers

Nospeciccellcycleanalysis sotware required


or multiparametric cell cycle research.

FlowCellect Bivariate Cell Cycle Kit

or DNA Replication Analysis

Investigate DNA replication in the S phase with high

accuracy and confdence. The kit includes a directly

conjugated Anti-BrdU Alexa Fluor 488 antibody plus

a DNA dye (propidium iodide). BrdU incorporation is awidely accepted method o measuring DNA replication

and kinetics o cell cycle progression. The percentage o

BrdU labeled cells is a reliable estimate o the S phase

compartment, and labeled cells can then be ollowed

through the cell cycle.

Detection o DNA replication

by analysis o S phase cells.

Bivariate ow cytometric

analysis using BrdU Alexa

Fluor 488 conjugate can

distinguish S phase cells

with great accuracy, not only

based on their dierencein DNA content rom G1 or

G2/M cells but also as having

incorporated BrdU.

G=24%

(-BrdU, 1X DNA content)

S=72%

(BrdU, 1-2X DNA content)

G2/M=4%

(-Brdu, 2X DNA content)

FEATuRED PRODuCT

Cell Growth

Cell Prepares

for Division

DNA Synthesis

Mitosis

Cells that

cease dividing

INTERPHAS

E

Cy

tokinesis

Propidium Iodide(x1000)

BrdU

104

103

102

101

1000 1 3 5 7 9

S Phase

G1 G2/M


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FlowCellect Bivariate Cell Cycle Kit

or G2/M Analysis

Investigate the G2/M phase transition with high accuracy

and confdence. The phosphorylation o histone H3 at

Ser10correlateswiththeG2toMphasetransitionandis

a prerequisite or chromatin condensation at mitosis. At

the end o mitosis, histone H3 is rapidly dephosphorylated

and remains unphosphorylated throughout the remainder

ofinterphase.Therefore,phospho-histoneH3(Ser10)isa

reliable, specifc marker o M-phase cells.

Nocodazole treatment increases percentage o cells in M phase.

Cell were either treated with 100 m Nocodazole (test sample)

or let untreated (control) overnight at 37 C. By plotting the

phosohorylation o histone H3 at Ser10 (y axis) versus DNA

content (x axis), an increase in the proportion o G2/M cells

was observed indicating that mitotic cells have accumulated

ater treatment. Apporomately 2% o cells reside in M phaseunder normal conditions in Jurkat cells, but when treated cell

population increased 18%.

PE (x1000)

Propidium Iodide

Control (untreated) Nocodazole treated

pH3-Alexa

Fluor488

pH3

104

103

102

101

1000 1 3 5 7 9

PE (x1000)

pH3-Alexa

Fluor488

104

103

102

101

1000 1 3 5 7 9

3% cellsin M phase

18% cellsin M phase

gava Cell Cycle Assay

This kit uses the nuclear DNA stain, propidum iodide (PI), to

measurecellcycle.Restingcells(G0/G1)containtwocopies

o each chromosome. Cycling cells synthesize chromosomal

DNA (S phase), which results in increased uorescence

intensity. When all chromosomal DNA has doubled (G2/M

phase), cells uoresce with twice the intensity o the initial

population.


Bivariate Cell Cycle Kit or DNA Replication Analysis Anti-BrdU / Propidium Iodide Solution 25 tests FCCH025102

BivariateCellCycleKitforG2/MAnalysisAnti-phospho-HistoneH3(Ser10)/PropidiumIodideSolution 25 tests FCCH025103

guava Cell Cycle Reagent Propidium Iodide Solution 100tests 4500-0220

Cell Cycle Phases:

G1 = 57%

S = 19%

G2 = 15%

M = 3%

FlowCellect Cell Cycle Kits

P1 (x1000)

pH3-Alexa

Fluor4

88

104

103

102

101

1000 1 3 5 7 9

M

G2

SG1

FlowCellect Bivariate Cell Cycle Kit or G2/M

Analysis (Application)

FEATuRED PRODuCT

Discrimination between G2 and M phase cells by measuring

the phosphorylation o histone H3 on Ser10. Histone H3 is

constitutively phosphorylated at Ser10 during metaphase.


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FEATuRED PRODuCTDNA DamageSignaling PathwayInvestigating the DNA damage signaling pathway is an

important area or genome health and cancer research.

Evidence suggests there is a direct correlation between

DNA damage and cell cycle. Cells that are deective in

DNA damage pathways can cause cancer because they

lack the ability to sense and repair the damage, leading to

genetic instability and ultimately uncontrolled cell growth.

The main kinase activated in response to double-stranded

DNA breaks is ATM or Ataxia telangiectasia mutated

kinase. ATM is a member o the phospho inositide 3-kinase

(PI3K)-related Ser/Thr protein kinase amily. Inactive ATM

exists as a dimer but quickly dissociates and becomes

phosphorylatedonSerine1981inresponsetoionizing

radiation. Once activated, ATM phosphorylates a number

o downstream actors, including P53, CHK2, SMC1, NBS1,and Histone H2A.X.


Mlticolor DNA Damage Response KitAnti-p-SMC1(S957)-AlexaFluor488/Anti-pATM(S1981)-PE/Anti-pHistoneH2A.X(S139)-PerCP

25 Tests FCCH025104

DNA Damage Histone H2A.X Dal Detection KitAnti-pHistoneH2A.X(Ser139)-PerCP/Anti-HistoneH2A.X-FITC

25 tests FCCS025153

Cell Cycle Checkpoint H2A.X DNA Damage KitAnti-pHistoneH2A.X(Ser139),cloneJBW301-AlexaFluor488/PropidiumIodideSolution

25 tests FCCH025142

Cell Cycle Checkpoint ATM DNA Damage KitAnti-pATM(Ser1981),clone10H11.E12-AlexaFluor488/PropidiumIodiden

25 tests FCCH025143

FlowCellect DNA Damage Histone H2A.X

Dal Detection Kit

FlowCellect Histone H2A.X DNA Damage Dual Detection kit

includes two directly conjugated antibodies, a phospho-

specicAnti-phospho-HistoneH2A.X(Ser139)-PerCP

and an Anti-Histone H2A.X-FITC conjugated antibody to

measure total levels o Histone H2A.X. This two color kit is

designed to detect the extent o Histone H2A.X pathway

activation by measuring H2A.X phosphorylation relative to

the total H2A.X expression in any given cell population. By

doing such, the levels o both the total and phosphorylated

protein can be measured simultaneously in the same cell,

resulting in a normalized and accurate measurement o

H2A.X activation ater stimulation. Moreover, simultaneous

measurement o both total and phospho-Histone H2A.X

confrms target specifcity o the phosphorylation event.

Together, a total and phospho antibody duo perormedin multiplex provides an enhanced and more reliable

detection o the phospho: total ratio within a mixed cell

population. Using this antibody pair provides a sensitive

and valuable tool to study the actors that induce DNA

damage and/or aect DNA repair, and allow one to explore

the linkage between DNA damage, cell cycle checkpoints,

and initiation o apoptosis.

FlowCellect DNA Damage Kits

p53

P

P

MRN

Complex

ATM

ATM

ATM

Ionizing Radiation Changes in

Chromatin Structure

BRCA1

CHK2

P

H2AX

PP

P

P

P

P

P

P

53BP1 MDC1

SMC1

P

P

Apoptosis

DNA RepairCell-cycle

Checkpoint

Arrest

P

P

A. Unstimulated B. Stimulated

H2AX-FTC (GRN-HLog)

Total H2AX - FITC

Phospho

H2AX

-

PerCP

pH2AX-PerCP

(RED-H_

cg)

H2AX-FTC (GRN-HLog)

pH2AX-PerCP

(RED-H_

cg)

Data below: Unstimulated

HeLa cells are stained with

both phospho-Histone

H2A.X-PerCP and Anti-Histone

H2A.X-FITC (A), where there isno indication o Histone H2A.X

activation via phosphorylation,

but only on total H2A.X as

noted by 97.2% o cells.

However, once HeLa cells

were stimulated with 100

M etoposide, simultaneous

measurement o both total

and phospho Histone H2A.X

confrms target specifcity o

the phosphorylation event

as indicated by the double

positive cell population (B)

as indicated by the 2.09%

to 97.15% increase o

double positive staining. Thelevels o both the total and

phosphorylated protein can

be measured simultaneously

in the same cell, resulting in

a normalized and accurate

measurement o H2A.X

activation ater stimulation.


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Mitochondrial AnalysisMitochondria are critical cellular organelles that produce

90%ofcellularenergy,controlcellsurvivalbyregulating

apoptosis, and produce reactive oxygen species (ROS).

Mitochondrial superoxide generation results in oxidative

stress, damage and cell death by apoptosis or to cellular

energetic decline. Thereore, mitochondrial dysunction

caused by disease or compound treatment has dire

consequences that can result in cell death.

Monitoring impact on mitochondria and related cell health

markers is an important part o drug screening programs,

pathway mapping, apoptosis, and disease research.

Flow cytometry detects multiple markers simultaneously at

various stages o apoptosis, making it a powerul technique

or studying pathways governing cell health and cell death.

FlowCellect Mitochondrial Kits

These kits harness the power o ow cytometry to assess

changes in mitochondrial membrane potential, apoptosis

as measured by Annexin V binding, mitochondrial oxidative

stress, and cell death, using only minimal cellular samples.

The kits may be used with most dual laser ow cytometry

systemsequippedwitha488nmanda644nmlaser.

Advantages

Assay:

Multiplexdetectionwithnooptimizationrequired

Highlyreproducible

Minimalassaydevelopmentneeded

Alloptimizedowcytometryantibodiesandbuffersincluded

Enablesnoviceuserstoperformcomplexanalysis

Samples:

Designedtorun100samplesofhumancells


or multiparametric mitochondrial

analysis.

Pro-Caspase 8

Pro-Caspase 9

Cyt c

Cyt c

APAF-1

Smac/Diablo

AIF

Endo G

Caspase 8

Caspase 3

Apoptosis

Mitochondria

Apo

ptosome

Nucleus

m MitochondrialPotential Change

Activated Caspase Cascade

ER Stress,

DNA Damage,Oxidants

DNA Fragmentation

Chromatin Condensation

Bax

Bak

t-Bid

IAP

IAP

Bid

Mitochondrial

Signaling andApoptosis

Adapted rom Bayir

and Kagan

Critical Care 2008

12:206.


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FlowCellect MitoDamage Kit

These kits harness the power o ow cytometry to assess

changes in mitochondrial membrane potential, apoptosis

as measured by Annexin V binding, mitochondrial oxidative

stress, and cell death, using only minimal cellular samples.

The kits may be used with most dual laser ow cytometry

systemsequippedwitha488nmanda644nmlaser.

Simltaneosly measres 3 important cell healthparameters:

Changeinmitochondrialpotential(consideredanearly

hallmark o apoptosis or cell stress)

Phospatidylserinetranslocationtothesurfaceofearly

apoptotic cells (measured by Annexin V binding)

Plasmamembranepermeabilizationorcelldeath

(measured by 7-AAD)

The FlowCellect MitoDamage kit can ths distingishmltiple poplations:

1. Healthy cells with intact mitochondrial membrane

2. Stressed cells with dissipated membrane potential

without Annexin V or 7-AAD staining

3. Early apoptotic cells with dissipated membrane

potential and Annexin V binding

4. Late apoptotic cells or dead cells with dissipated

membrane potentials

Uninduced 50 M CCCP

MitoSenseRed

Annexin V, CF488A

Red2Fluoresecence(RD2-HLog)

Green Fluorescence (GRN-HLog)

94.4%

0.75% 3.7%100 101 102 103 104

100

101

102

103

104

1.1%



54.7%

14.6% 27.0%100 101 102 103 104

100

101

102

103

104

3.7%



0.20%

93.2% 6.6%100 101 102 103 104

100

101

102

103

104

0.04%

MitoSenseRed

Red2Fluorescence(RD2-HLog)

Red Fluorescence (RED-HLog)

95.2%

3.2% 1.3%100 101 102 103 104

100

101

102

103

104

0.3%


Red Fluorescence (RED-HLog)

0.26%

98.4% 1.3%100 101 102 103 104

100

101

102

103

104

0.02%

100 101 102 103 104

101

102

103

104

Depolarized Cells

Dead Cells

Live Cells

58.1% 0.08%

41.0% 0.8%


Red Fluoresecence (RED-HLog)

100

2 M Staurosporine

7-AAD

Annexin V, CF488A



0.16%

95.2% 3.2%100 101 102 103 104

100

101

102

103

104

1.4%



0.06%

70.5% 28.2%100 101 102 103 104

100

101

102

103

104

1.2%



0.10%

93.9% 4.8%100 101 102 103 104

100

101

102

103

104

1.2%

7-AAD

Dot plots depicting Jurkat cells

stained using the MitoDamage

kit. Jurkat cells uninduced,

induced to apoptosis with

2 M staurosporine or with

50 M CCCP, then stained

using the MitoDamage kit.

Plots show the percentage o

positive cells or:

1st row: Apoptosis (AnnexinV binding) and mitochondrial

membrane potential change

2nd row: Cell death and

mitochondrial membrane

potential change

3rd row: Apoptosis and cell

death.

Data reports that 2 M

staurosporine induces

apoptosis in Jurkat cells, and

that 50 M CCCP depolarizes

the mitochondrial membrane,

but neither condition issufcient or cell membrane

permeabilization and death.

Kit Component Laser Ber Pack

MitoSense Red Dye Red10XAssay

Buer HSCAnnexin V-CF488A Blue

7-AAD Blue

FEATuRED PRODuCT


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FlowCellect MitoPotential Red Kit

Simultaneous analysis o mitochondrial membrane

potential along with cell death provides key inormation

or drug discovery, cancer and toxicology studies as well

as disease-induced apoptosis. This kit uses MitoSense

Red (a red laser-excitable dye) to monitor mitochondrial

membrane potential changes in early apoptosis, and

7-AAD (a live-cell-impermeant DNA intercalator) tosimultaneously monitor cell membrane permeability

changes in late apoptosis and necrotic cell death.

FlowCellect MitoLive Kit

Incldes:

MitoSense Red, a uorescent cationic dye that

accumulates in the mitochondria and is responsive to

mitochondrial potential changes

Calcein acetoxymethylester (calcein-AM) a non-

uorescent, cell-permeant compound that is hydrolyzed

by intracellular esterases into the uorescent anion

calcein and provides a measure o cellular vitality.

The simultaneous use o the reagents enables researchers

to obtain inormation on early and late apoptosis in one

simple assay.

FlowCellect MitoStress Kit

Incldes:

MitoSOX Red, a live-cell-permeant, uorogenic

dye which targets the mitochondria and reacts with

superoxide radicals and uoresces yellow/red

AnnexinVconjugatedtoCF647whichbindsto

phosphatidylserine (PS) on the surace o apoptotic cells

The simultaneous use o these reagents enables researchers

to obtain inormation on oxidative stress and apoptosis in

one simple assay.

FlowCellect Cytochrome c Kit

Includes a directly labeled anti-Cytocrome cFITC antibody

and Anti-IgG1-FITC isotype control. Viable or live cells will

demonstrate higher levels o Cytochrome c staining while

apoptotic cells which have released their Cytochrome c

rom the mitochondria to the cytoplasm will demonstrate

reduced staining intensity. The FlowCellect Cytochrome c

ow cytometry kit is a simple, gentle, and ast method to

assess levels o Cytochrome c in mitochondria, providing a

valuable tool or assessing pro-apoptotic signaling and the

efcacy o pro-apoptotic anti-cancer agents in cells.


FlowCellect MitoPotential Red Kit

Two dyes or measuring cell death and mitochondrial membrane potential MitoSense Red (Red Laser) / 7-AAD (Blue Laser)

100tests FCCH100105

FlowCellect MitoDamage Kit

Three dyes to assess mitochondrial potential, stress, and cell death

Mitosense Red (Red) / Annexin V-CF488A (Blue) / 7-AAD (Blue Laser)

100tests FCCH100106

FlowCellect MitoLive Kit

Two dyes to measure mitochondrial health and cell vitality

Mitosense Red (Red) / Calcein-AM (Blue Laser)

100tests FCCH100107

FlowCellect MitoStress Kit

Understanding the regulation o apoptosis and oxidative stress.

MitoSoxRed(Red)/AnnexinV-CF647(BlueLaser)

100tests FCCH100109

FlowCellect Cytochrome c Kit

Easy way to detect loss o mitochondrial Cytochrome c in cells

Anti-Cytochrome c-FITC (Blue) / Anti-IgG1-FITC (Blue Laser)

100tests FCCH100110

FlowCellect Oxidative Stress Characterization Kit

Detection o oxidative stress by ow cytometry

Anti-DNP-FITC (Blue Laser)

25 tests FCCH025111

guava Mitochondrial Depolarization Assay Kit

Monitoring changes in mitochondrial membrane potential

JC-1 (Blue Laser) / 7-AAD (Blue Laser)

100tests 4500-0250

FlowCellect Mitochondrial Kits


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Early Mid Late

Guava Nexin Multicaspase Guava TUNEL

Annexin Red Caspase 3/7, 8, 9 Assay

MitoPotential Red Dual Caspase

MitoDamage

MitoStress

MitoLive

Cytochrome c

c

cc

c

c+

PS

PS

PS

PS

PS

PS

c+c+

c+

c+

c+

G...C

G...C

C...G

A...T

G...C

C...G

A...T

A

G

G...C

G...C

C...G

A...T

G...C

C...G

A...T

A

G

G...C

G...C

C...G

A...T

G...C

C...G

A...T

A

G

TdT

+

BrdUTP

TRITC-

Anti-BrdU

DNA strand

breaks de to

apoptosis

Add BrduTP

to 3OH ends

Antibody labeled

break sites

ApoptosisCells respond to specifc apoptotic signals by initiating

intracellular processes that result in characteristic

physiological changes. Among these changes areexternalization o phosphatidylserine to the cell surace,

depolarization o mitochondrial membranes, cleavage

and degradation o specifc cellular proteins, compaction

and ragmentation o nuclear chromatin, loss o cell

membrane integrity, and cellular shrinkage. Suppression or

enhancement o apoptosis is known to cause or contribute

to diseases such as cancer and diabetes, making the

apoptotic pathway a popular drug target.

Because apoptosis is a dynamic event, and the time period

during which cells exhibit apoptosis markers is variable andshort, ow cytometry is an ideal technique or tracking

cells through apoptosis. Our easy-to-use kits enable you to

examine cells at each o the various stages o apoptosis.

Early Apoptosis Flow Cytometry Kits

Two separate dyes identiy a broad spectrum o

apoptotic and non-apoptotic cells: Annexin V binds to

phosphatidylserine on the external membrane o apoptotic

cells, while 7-AAD permeates and stains DNA o late-stage

apoptotic and dead cells.

Advantages

Twodyestrategy:Detectvariousstagesofapoptosiswithin a one assay

Mix-and-readassay:Getstandardizedresultsevenwithmultiple users

All-in-one-kit:Spendlesstimebeforeanalysis

CompatiblewithpairingwithFITCorPEprobesorotherprobes in the green or yellow channels

Probewithothermarkersingreenandyellowchannels

with the FlowCellect Mitochondrial kits

Mid-Apoptosis Flow Cytometry Kits

Caspase activity is measured using a FLICA (uorescent

labeled inhibitor o caspase) reagent, supplemented by

a nuclear DNA stain 7-AAD, which evaluates membraneintegrity and cell viability. The assays are available in two

orms, sulorhodamine (SR) and carboxyuorescein (FAM),

giving greater exibility in assay design as well as the

capacity to multiplex caspase assays.

Late Apoptosis Flow Cytometry Kits

The guava TUNEL assay detects apoptosis-induced DNA

ragmentation through a quantitative uorescence assay.

Terminal deoxynucleotidyl transerase (TdT) catalyzes the

incorporation o bromo-deoxyuridine (BrdU) residues into

the ragmented nuclear DNA at the 3-hydroxyl ends. A

TRITC-conjugated anti-BrdU antibody then labels these

DNA ragments. The assay distinguishes two populations:

non-apoptotic cells (TUNEL-negative) and apoptotic cells

(TUNEL-positive).

Discover the power o ow cytometryor multiparametric apoptosis analysis.

Live/Healthy

(non-committed)cells

FLICA(-)7-AAD(-) FLICA(+)7-AAD(-) FLICA(+)7-AAD(+) FLICA(-)7-AAD(+)

Early/Mid-stage

(committed)apoptotic

cells

Late stage

apoptotic/dyingcells

Dead cells

C+C+C+

C+

C+C+ C+

C+PI+PI+

PI+

PI+PI+

PI+


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FlowCellect Annexin Red Kit

In the early stages o apoptosis, phosphatidylserine

molecules, which can bind to Annexin V, move rom the

inner leaet, to the outer leaet o the plasma membrane.

A rapid, sensitive, and convenient assay to monitor early

and late apoptosis, the FlowCellect annexin red kit includes

recombinant Annexin V conjugated to the red sensitive

dyeCF647,and7-AAD(alivecell-impermeantdye)to

measure cell membrane integrity. Ater staining cells with

FlowCellect Annexin Red kit, three populations o cells can

be identifed in this assay:

Non-apoptotic cells: Annexin V(-) and 7-AAD(-)

Early apoptotic cells: Annexin V(+) and 7-AAD(-)

Late-apoptotic or dead cells: Annexin V (+) and 7-AAD(+)

Two Dyes to distinguish early Apoptosis rom later stages

Dot plots depicting Jurkat cells stained using the FlowCellect Annexin Red kit. Jurkat cells were untreated (Plot A), treated with0.1 M (Plot B) or with 3 M staurosporine (Plot C), and then stained using the FlowCelllect Annexin Red kit. The percentage o

apoptotic cells increased rom 36% to 92% in response to a 30-old increase in staurosporine concentration; however, only a small

raction (< 10%) o the cells showed evidence o cell death.

Kit Component Laser Ber Pack

AnnexinV-CF647 Red 10XAssay

Buer HSC7-AAD Blue

FEATuRED PRODuCT

7-AAD

B. C.

Annexin V

A.


2% 2%

85% 11%

Red2 Fluorescence (RED2-HLog)

100 101 102 103 104100

101

102

103

104

100

101

102

103

104


4%

60% 36%


100 101 102 103 104100

101

102

103

104


7%

5% 92%


100 101 102 103 104

3 M Staurosporine0.1 M StaurosporineUninduced

PS = phosphatidylserine

Apoptotic Cell MembranesHealthy Cell Membranes

Annexin CF647

PSPS

PS

PS

PSPS

PS

PS

= 7aad

FlowCellectAnnexinRedKit(AnnexinV-CF647Reagent/7-AAD) 100tests FCCH100108

Guava Nexin Reagent (Annexin V-PE/ 7-AAD) 100tests 4500-0450

Guava TUNEL Reagent Kit (Anti-BrdU -TRITC/ TdT Enzyme/ Br-dUTP) 100tests 4500-0121

FlowCellectBcl-2ActivationDualDetectionKit(Anti-Bcl-2-AlexaFluor488/Anti-pBcl-2(Ser70)-PE) 25 Tests FCCS025108

Guava MultiCaspase SR Kit (MultiCaspase SR reagent / 7-AAD) 100tests 4500-0500

Guava Caspase 3/7 SR Kit (Caspase 3/7 SR reagent / 7-AAD) 100tests 4500-0510

Guava MultiCaspase FAM Kit (Multicaspase FAM reagent / 7-AAD) 100tests 4500-0530

Guava Caspase 3/7 FAM Kit (Caspase 3/7 FAM reagent / 7-AAD) 100tests 4500-0540

Early Apoptosis Kits

Late Apoptosis Kit

Apoptosis Signaling Kit

Mid Apoptosis Kits

For a complete listing visit: www.millipore.com/midapoptosis_kits

Apoptosis Kits



13/32

13


FlowCellect EGFR/MAPK Pathway Activation Detection KitAnti-pEGFR (Tyr1173), AlexaFluor 488

Anti-pERK1/2(Thr202/Tyr204,Thr185/Tyr187)-PE

25 Tests FCCS025101

FlowCellect EGFR RTK Activation Dal Detection KitAnti-pEGFR (Tyr1173), AlexaFluor 488

Anti-EGFR-PerCP

25 Tests FCCS025107

FlowCellect EGFR/STAT3 Pathway Activation Detection KitAnti-pEGFR (Tyr1173), Alexa Fluor 488

Anti-pSTAT3(Tyr705),AlexaFluor647

25 Tests FCCS025111

FlowCellect Kits: MAPK Pathway

FlowCellect Kits: EGFR Pathway

Cell Signaling

Signal transduction pathways lead to diverse outcomes,

such as apoptosis, cell dierentiation, cell growth and cell

prolieration, all o which have been extensively studied

in the process o developing therapies or various cancers

and autoimmune disease. Cross-talk among signaling

pathways adds an extra dimension o complexity when

analyzing physiological consequences o a pathway o

interest. However, there are some key nodes at which

multiple signals are integrated. Multiparametric ow

cytometry provides researchers the power to monitor these

intracellular checkpoints simultaneously, enabling analysis

o complicated cell events.

FlowCellect Cell Signaling KitsThe study o cell signaling has been made easier by

activation status-specifc and phospho-specifc antibodies.

Measuring the activity o cell signaling pathways by ow

cytometry delivers robust, high content inormation in

less time than traditional methods by analyzing multiple

parameters on hundreds o cells per second.

Dal Detection FlowCellect Kits With Pairs o

Total and Phospho-Specic Antibodies

Merck Millipores FlowCellect Dual Detection kits are

a series o ow cytometry products which include a

pair o antibodies that bind to the same protein; one to

detect total protein expression and another to detect

the phosphorylated orm o the same target. Using

two parameter analysis, we can achieve target specifc

detection o phosphorylation and, by doing so, eliminate

alse positives while enhancing the signal to noise ratio.

FlowCellect Cell Signaling Kits With Directly

Conjgated, Phospho-Specic Antibodies

Determine the eect o mechanical and chemical reagents

that can induce DNA damage, discern multiple pathway

activation and cross talk in a time-dependent manner,

or study the correlation between pathway activation and

changes in cell unction and health.


or multiparametric cell signaling research.


FlowCellect PI3K/MAPK Dal Pathway Activation and Cancer Marker Detection KitAnti-pErk1/2(Thr202/Tyr204,Thr185/Tyr187)-PE

Anti-phospho-Akt1/PKBa (Ser473), Alexa Fluor 488

Anti-KI-67-PerCP

25 Tests FCCS025100

FlowCellect EGFR/MAPK Pathway Activation Detection KitAnti-pEGFR (Tyr1173), AlexaFluor 488

Anti-pERK1/2(Thr202/Tyr204,Thr185/Tyr187)-PE

25 Tests FCCS025101

FlowCellect MAPK Activation Dal Detection KitAnti-pERK1/2(Thr202/Tyr204,Thr185/Tyr187)-PE

Anti-ERK1/2-AlexaFluor647

25 Tests FCCS025106

FlowCellect p38 Stress Pathway Activation Detection KitAnti-pP38(Thr180/Tyr182),AlexaFluor488

Anti-pATF2(TThr69/71),AlexaFluor647

25 Tests FCCS025132

Advantages

Assay:

Ensuresspeciclabelingo targets

Multiplexdetectionwithno optimization required

Enablesnoviceusersto perorm complexanalysis

Samples:

Designedtorun25samples o human cells


14/32

14

FlowCellect PI3K/MAPK Dal Pathway

Activation and Cancer Marker

Detection Kit

Three antibodies to study cross-talk between the PI3K and

MAPK pathways. The kit uses directly labeled antibodies

against phospho-Akt1/PKBa(Ser473)-AlexaFluor 488 and

Anti-phospho-Erk1/2(Thr202/Tyr204,Thr185/Tyr187)-PE

toanalyzesignalingactivationandcrosstalk,plusKi-67

marker-PerCP marker to identiy the prolierative raction.

Together, the antibody trio makes it easy to evaluate the

role these signaling pathways play in prolieration and

dierentiation. The kit also includes Cell Cycle Stop

xationreagenttoimproveKi-67detection.Although

Ki-67ispresentthroughoutmostofthecellcycle,itis

difcult to detect by ow cytometry except during M

phase. Cell cycle stop reagent arrests the cycle at the

Mphase,makingitpossibletoaccuratelydetectKi-67

expression and discern the biological eects o the PI3K/

MAPK cross talk.

FEATuRED PRODuCT

GTP

GDP

Shc

Grb2

Ki 67

PI3-Kinase

Proliferation/

Survival

Growth/

Differentiation

Wortmannin

or LY294002

PMA

p85

IGF-1

p110 RAS

Raf-1Akt

P

P

P

P

P

SOS

MEK1/2

Inhibiting

Activating

ERK1/2

P

The cross-talk between the PI3K/Akt and MAPK signaling pathways is evident downstream o IGF in HEK293 cells. Cells are

stimulated at 3 minutes and 5 minutes by Insulin and PMA as shown in A and B, respectively. As previously indicated, although

Insulin initially activates both pathways independently, the activation o the PI3K pathway indicated by the phosphorylation o Akt

and ERK1/2 inhibits the activation o ERK. The dot plots illustrate the cross-talk that exists between these two signaling pathways.

This is demonstrated by the sharp decrease in ERK phosphorylation between 3 minutes and 5 minutes (see bar graphs; C, D).

pAKT-Alexa 488

pAKT

Untreated

A. 3 Minute Stimulation C. HEK203 - 3 Minute Stimulation

D. HEK203 - 5 Minute StimulationB. 5 Minute Stimulation

Insulin Treated PMA Treated

pERK-PE

pERK

104

104

103

103

102

102

101

101100

100

pAKT-Alexa 488 pAkt pErk

Untreated100 nM insulin 3 min100 g/mL PMA 3 min

Ki67

010

pERK-PE

%o

fcells

104

104

103

103

102

102

101

101100

100

pAKT-Alexa 488

pERK-PE

104

104

103

103

102

102

101

101100

100

pAKT-Alexa 488

pERK-PE

104

104

103

103

102

102

101

101100

100

pAKT-Alexa 488

pERK-PE

104

104

103

103

102

102

101

101100

100

pAKT-Alexa 488

pERK-PE

104

104

103

103

102

102

101

101100

100

2030405060708090

100

pAkt pErk

Untreated100 nM insulin 3 min

100 g/mL PMA 3 min

Ki67

010

%o

fce

lls

2030405060708090

100


15/32

15

FlowCellect Kit EGFR RTK Activation

Dal Detection Kit

EGF pathway activation through EGFR plays a key role

in the regulation o essential cellular processes, and

abnormality o EGF signaling is linked to cancer. Merck

Millipores FlowCellect EGFR RTK Activation Dual Detection

kit includes two directly conjugated antibodies, a phospho-

specifc Anti-phospho-EGFR (Tyr1173)-Alexa Fluor 488

and an Anti-EGFR-PerCP conjugated antibody to measure

total levels o EGFR. This two color ow cytometry kit is

designed to detect the extent o EGF pathway activation

by measuring the EGFR phosphorylation in relative to the

total EGFR expression in any given cell population. By

doing such, the levels o both the total and phosphorylated

protein can be measured simultaneously in the same cell,

resulting in a normalized and accurate measurement o

EGFR activation ater stimulation. Moreover, simultaneous

measurement o both total and phospho-EGFR confrms

target specifcity o the phosphorylation event. Together,

a total and phospho antibody duo perormed in multiplex

provides an enhanced and more reliable detection o the

phospho:total ratio within a mixed population.

FEATuRED PRODuCT

EGF

EGFR

Ras

ERK

PI3KJAK

AKTSTAT

P P

mTOR

EGFR-PerCP

Total EGFR

A. Isotype Control(No EGF Stimulation)

B. Total and pEGFR(No EGF Stimulation)

C. Isotype Control(EGF Stimulation)

D. Total and pEGFR(EGF Stimulation)

pEGFR-Alexa

Fluor488

pEGFR

104

104

103

103

102

102

101

101100

100

EGFR-PerCP

pEGFR-Alexa

Fluor488

104

104

103

103

102

102

101

101100

100

EGFR-PerCP

pEGFR-Alexa

Fluor

488

104

104

103

103

102

102

101

101100

100

EGFR-PerCP

pEGFR-Alexa

Fluor48

8

104

104

10

3

103

102

102

101

101100

100

98% 88.6%

5.4%0.5%

5.5%

1.0%

0.6%

0.7%

0% 0.3%

0.5%

0% 1.5%

0.2%

98.2

99%

Dual Parameter Analysis o

Total and Phospho EGFR on

A431Cells As illustrated indot plots A and B, untreated

A431cells stained with an

isotype control (A) and both

pEGFR-Alexa Fluor 488

and Anti-EGFR-PerCP (B),

respectively, only indicated

EGFR expression on total EGFR

as noted by 89% o cells.

However, once A431cells were

stimulated with 100 ng/

mL EGF, simultaneous

measurement o both total

and phospho EGFR confrms

target specifcity o the

phosphorylation event as

indicated by the doublepositive cell population (D) as

indicated by the 5% to 98%

increase o double positive

staining. A431 stimulated

cells showed no activity

when stained with an isotype

control (C).


16/32

16

FlowCellect Chemokine Receptor

Srace Expression Qantication KitsMerck Millipore oers 11 GPCR surace identifcation

ow cytometry kits. Flow cytometry provides high quality,

reproducible data in ar less time than traditional methods

or chemokine receptor research and avoids the hazard and

expense o radioligand binding assays.

Our FlowCellect GPCR identifcation kits can be used to

identiy and quantiy GPCRs on the surace o any cells.

The kits detect GPCR expression using a GPCR-specifc

antibody validated or ow cytometry. Also included are

positive and negative control cells with well-characterized

receptor expression levels or the purposes o quantitative

extrapolation.

Advantages

Assay:

Includespharmacologicallycharacterizedpositiveandnegative control cells

Sameaccuracyasradioactiveassays,withoutthehazards.

Abilitytoidentifyhigh,medium,andlowexpressingcellcultures during the clonal selection process.

HighreproducibleresultsobtainedbyusingMerckMillipores well-characterized ChemiScreen GPCR celllines as assay controls

Samples: Sufcientreagentstorun100samplesofhumancells.

For a complete listing o kits visit: millipore.com/fowcytometry.


FlowCellectChemokineReceptorCCR2BSurfaceExpressionQuanticationKit 100tests FCCR200411

FlowCellectChemokineReceptorCCR4SurfaceExpressionQuanticationKit 100tests FCCR400413

ChemokineReceptorCCR6SurfaceExpressionQuanticationKit 100tests FCCR600414

FlowCellectChemokineReceptorCCR7SurfaceExpressionQuanticationKit 100tests FCCR700415

FlowCellect PI3K/MAPK Dal Pathway Activation and Cancer Marker Detection KitAnti-phospho-Erk1/2(Thr202/Tyr204,Thr185/Tyr187)-PE/Anti-p-Akt1/PKBa (Ser473)- Alexa Fluor 488 /

Anti-KI-67-PerCP

25 Tests FCCS025100

FlowCellect PI3K Activation Dal Detection KitAnti-phospho-Akt(Ser473)AlexaFluor488/Anti-Akt/PKB-AlexaFluor647

25 Tests FCCS025105

FlowCellect PI3K-mTOR Signaling Cascade Mapping KitAnti-p-RibosomalProteinS6(Ser235)-PerCP/Anti-p-Akt1/PKBa (Ser473)-Alexa Fluor 488

25 Tests FCCS025210

FlowCellect EGFR/STAT3 Pathway Activation Detection KitAnti-pEGFR(Tyr1173),AlexaFluor488/Anti-pSTAT3(Tyr705),AlexaFluor647

25 Tests FCCS025111

FlowCellect Mlti-STAT Activation Proling KitAnti-pSTAT1(Tyr701)-PerCP/Anti-pSTAT3(Tyr705)-Alexa488/Anti-pSTAT5A/5B(Tyr694/Tyr699)-PE

25 Tests FCCS025550

FlowCellect STAT1 Activation Dal Detection KitAnti-p-STAT1(Tyr701)AlexaFluor488/Anti-STAT1-PerCP

25 Tests FCCS025142

FlowCellect STAT3 Activation Dal Detection KitAnti-p-STAT3(Tyr705)AlexaFluor647/Anti-STAT3-AlexaFluor488

25 Tests FCCS025143

FlowCellect Bcl-2 Activation Dal Detection KitAnti-Bcl-2 Alexa Fluor 488 Antibody

Anti-pBcl-2(Ser70)PE

25 Tests FCCS025108

Chemokine Receptor Kits

PI3/ Akt/ m-TOR Pathway

Jak/ STAT Pathway

Mltiple Pathway

Apoptosis Signaling Pathway


FlowCellect Cell Signaling Kits

FlowCellect Src Activation Dal Detection KitAnti-pSrc(Tyr416)-AlexaFluor488

Anti-Src-AlexaFluor647

25 Tests FCCS025154

FlowCellect PLC-1 Activation Dal Detection KitAnti-pPLC-1 (Tyr783) Alexa Fluor 488

Anti-PLC-1-PE

25 Tests FCCS025145


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17

FlowCellect Hman ESC (Oct4) Nclear Marker Characterization Kit

hOCT4-Alexa 488 / SSEA4-PE / SSEA1-PE/CY5

25 Test FCHEC25102

FlowCellect Hman ESC (HESCA-1) Srace Marker Characterization KitHESCA1-FITC / SSEA4-PE / SSEA1-PE/CY5

25 Tests FCHEC25104

FlowCellect Hman ESC (TRA-1-60) Srace Marker Characterization KitTRA-1-60-FITC/SSEA4-PE/SSEA1-PE/CY5

25 Tests FCHEC25106

Hman

FlowCellect Rodent NSC Characterization Kit (Neronal Dierentiation)Sox-2 FITC / Nestin-PE / Beta-III-Tubulin-PE/CY5

25 Tests FCRNC25112

Rodent

FlowCellect Mose ESC (Oct4) Nclear Marker Characterization KitmOCT4-Alexa 488 / SSEA4-PE / SSEA1-PE/CY5

25 Tests FCMEC25110

Mose

Stem Cells

Because ow cytometry has the power to characterize

subpopulations o cells within heterogenous cell mixtures;

it is widely used or studying both embryonic stem cells

(ESCs) and induced pluripotent stem (iPS) cells. Flow

cytometry enables researchers to evaluate percentages

o cells expressing specifc markers, to determine culture

quality, and to track gene expression changes during a

dierentiation protocol.

FlowCellect Stem CellCharacterization KitsMerck Millipores FlowCellect stem cell characterization

kits are designed to provide rapid, sensitive assessments

o embryonic and neural stem cell phenotypes at various

stages. The kits use three parameters or accurate

identifcation, enabling the research to triangulate

cellular phenotypes with two complementary positive

markers and one negative marker. The negative antibody

also serves as a stage- and species-specifc or lineage-

specifc marker or dierentiated cells.

Advantages

Assay:

Alloptimized,uorescently-labeledowcytometryantibodies and buers included

Highlyreproducibleresults

Validatedforowcytometry,immunocytochemistry,andWestern blotting


or multiparametric stem cell analysis.

Embryonic Stem Cell Markers

Stem cell(hESC, mESC)

Oct-4+

SSEA-4+ (human)

TRA1-60+

HESCA+Nanog+

SSEA-1+ (mouse)

Committed cell(ENStem-A)

Oct-4-/+

SSEA-4-

TRA1-60-

HESCA-

DierentiatedcellsOct-4-

SSEA-4-

TRA1-60-

HESCA-

SSEA-1+/-


Stem Cell FlowCellect Kits


18/32

18

FlowCellect Hman Embryonic Stem Cell

Characterization Kit

The kit is provides a quick, easy way to track surace marker

expression o hOCT4, SSEA4 and SSEA1. The percentage o

undierentiated human embryonic stem cells in culture is

reected in the percentage cells that express both hOCT4

FlowCellect Hman Embryonic

Stem Cell HESCA-1 Srace Marker

Characterization Kit

This kit provides an easier and quick way to track surace

marker expression o HESCA-1, SSEA4 and SSEA1.

The kit will also help to determine the percentage o

undierentiated human embryonic stem cells in culture

by determining the percentage o cells that express both

and SSEA4 but not SSEA1. This quick test will enable

researchers to determine the multipotency o their cells in

culture as well as see changes in marker expression during

a dierentiation protocol.

HESCA-1 and SSEA4 and not SSEA1. This quick test will

enable researchers to test the quality o their cells inculture as well as see changes in marker expression during

a dierentiation protocol.

FEATuRED PRODuCTS

Representative data o H1 human embryonic stem cells stained with hOCT4-Alexa 488, SSFA4-PE and SSFA1-PF/CY5

Representative data o H1 human embryonic stem cells stained with HESCA-1-FITC, SSEA4-PE and SSEA1-PE/CY5.

0%

4.9%

0.1%

95%

10e4

10e3

10e2

10e1

10e0

10e0 10e1 10e2 10e3 10e4

SSEA1-PE/CY5

SSEA4-PE

10e4

10e3

10e2

10e1

10e0

10e0 10e1 10e2 10e3 10e4

HESCA-FITC

SSEA4-PE

10e485.5%

14.4%

0.1%

0%

10e3

10e2

10e1

10e0

10e0 10e1 10e2 10e3 10e4

hOCT4-Alexa488

SSEA1-PE/CY5

B. C.A.

B. C.A.

10e40.1%

4.8%

85.5%

9.6%

10e3

10e2

10e1

10e0

10e0 10e1 10e2 10e3 10e4

hOCT4-Alexa488

SSEA4-PE

10e4

10e3

10e2

10e1

10e0

10e0 10e1 10e2 10e3 10e4

HESCA-FITC

SSEA1-PE Cy5

10e4

10e3

10e2

10e1

10e0

10e0 10e1 10e2 10e3 10e4

SSEA1-PECy5

SSEA4-PE


19/32

19

Immunology

The immune system, which mediates the bodys response

to the introduction o oreign material, is made up

o multiple cell types collectively called lymphocytes.

Lymphocyte subtypes include B cells (which secrete

antibodies), cytotoxic T cells, helper T cells (which secrete

cytokines), and natural killer (NK) cells. Characterization o

lymphocyte subtypes and cytokine signaling is essential or

understanding the complex nature o the immune system.

Activation by antigens, suppression o normal immune

activation, and disease states can aect the phenotypes

o lymphocytes. Multiparameteric phenotypic analysis

by ow cytometry allows researchers to distinguish one

subpopulation o cells within a heterogeneous mixture,

and thereby enables the study the dynamics o immune

signaling in intact cells.

FlowCellect Immnology KitsEach kit includes multiple optimized uorescent labeled

antibodies and all buers necessary or cell preparation and

analysis. Detailed assay instructions are included to assist in

analysis and to ensure that the correct cell concentration is

obtained during acquisition o sample data.

Advantages

Assay:

Multiplexdetectionwithnooptimizationrequired

Highlyreproducible

Minimalassaydevelopmentneeded

Alloptimizedowcytometryantibodiesandbuffersincluded

Enablesnoviceuserstoperformcomplexanalysis

Samples:

Designedtorun25tests(FlowCellectkits)

Designedtorun100tests(guavakit)

Discover the power o ow cytometry ormultiparametric analysis o theimmune system.

DC

IL-12R

TH1

T-bet

STAT4

STAT1

IL-2R

TREG

Foxp3

STAT5

IL-21R

TFHTH

Bcl-6

STAT3

IL-4R

TH2

GATA3

STAT6

STAT5a

IL-23R

TH17

RORt

STAT3

IL-12

CD80

CD28

TCR

CD28

CD85

pMHCII

IFN

IL-18

IL-2

IL-4

IL-33

IL-6

IFN

IL-2

LT

IL-21

IL-6

TGF

IL-23

TGF

IL-2

IL-4

IL-5

IL-13

IL-25

IL-17

IL-17F

IL-22

IL-21

IL-21

IL-17

TGF

IL-10

IL-35

B ZONE

CD62Llo CCR7lo

CCR7lo

High

TCR Binding

Low

TCR Binding

Medium

TCR Binding

CXCR5hi

CD62Lhi CCR7hi T ZONE

EMIGRANTnaive

CD4+ T Cell Dierentiation. Schematic diagram o CD4+ T-cell dierentiation. Five dierent types o CD4+ T-cells can develop rom

a common nave precursor depending on the cytokine environment and interaction with dendritic cells (DC).


20/32

20

FlowCellect Mose Th1/Th17

Intracelllar Cytokine KitMerck Millipores FlowCellect Mouse Th1/Th17 Identifcation

Kit is designed to enable a researcher a quick and easy way

to detect IFN-g and IL-17 expression in mouse Th1 and

Th17 CD4+ T-cells. This kit contains an anti-IFN- antibody

conjugated to PE, an anti-IL17 antibody conjugated FITC

and an anti-CD4 antibody conjugated to PerCP. The kit also

includes optimized a protein transport inhibitor and buers

to aid in identiying Th1 and Th17 CD4+ T-cell subsets in ex

vivo lymphocyte populations or to monitor Th1 and Th17

CD4+ Tcell dierentiation in culture. In addition, Merck

Millipore has added a fxable viability dye that eliminates

alse positive data resulting rom cytokine staining on dead

cells that can occur in long term cultures. The kit contains

sufcient reagents or 25 3-color tests. Detailed assay

instructions are included to assist in sample preparation

and to ensure that the correct cell concentration is obtained

during acquisition o sample data. This kit is not designed to

be used in the analysis o whole spleen cells in SJL mice.

FEATuRED PRODuCT


FlowCellect Hman FOXP3 Treg Characterization KitAnti-CD3-PE/Cy5/Anti-CD4-FITC/Anti-FOXP3-AlexaFluor647

25 Tests FCIM025118

FlowCellect Hman CD4/CD8 T Cell KitAnti-CD8-FITC, CD4-PE, CD3-PECY5 o cocktail

100Tests FCIM100158


FlowCellect Mose Th1 Intracelllar Cytokine KitAnti-CD4 clone GK1.5-PerCP /Anti-IFN, clone XMG1.2-PE

A quick and easy way to detect IFN-expression in mouse Th1 CD4+ T cell

25 Tests FCIM025123

FlowCellect Mose Th2 Intracelllar Cytokine KitAnti-CD4-PerCP clone GK1.5/ Anti-IL-4, clone 11B11-PE

A quick and easy way to detect IL-4 expression in mouse TH2 CD4+ T-cells

25 Tests FCIM025124

FlowCellect Mose Th17 Intracelllar Cytokine KitAnti-CD4-PerCPcloneGK1.5/Anti-IL-17-,cloneTC11-18H10-FITC

A quick and easy way to detect IL-17 expression in mouse TH17 CD4+ T-cells.

25 Tests FCIM025125

FlowCellect Mose Th1/Th2 Intracelllar Cytokine Kit20XAnti-CD4cloneGK1.5-PerCP/Anti-IL-4,clone11B11-PE/Anti-IFN-, clone XMG1.2-PE

A quick and easy way to detect IFN- and IL-4 expression in mouse Th1 and Th2 CD4+ T-cells.

25 Tests FCIM025137

FlowCellect Mose Th1/Th17 Intracelllar Cytokine KitAnti-CD4-PerCPcloneGK1.5/Anti-IL-17,cloneTC11-18H10-FITC/Anti-IFN, clone XMG1.2-PE

A quick and easy way to detect IFN- and IL-17 expression in mouse TH1 and TH17 CD4+ T-cells.

25 Tests FCIM025138


FlowCellect Mose FOXP3 Treg Identication KitAnti-CD4cloneGK1.5-PerCP/Anti-FOXP3,clone3G3-AlexaFluor647

25 Tests FCIM025126

FlowCellect Mose Viable Treg Characterization KitAnti-CD4-PerCP/Cy5.5 / Anti-CD25-PE/ Anti-Foxp3-Alexa Flour 488

25 Tests FCIM025168

Hman

Mose

Mose

Reglatory T-cell Kits

Helper T-cell Kits

3.7% 56.0%

4.2% 36.1%

FVD-660

Th-1 Cells Th-17 Cells

Side

Scatte

r

7000

9000

5000

3000

1000

0

CD4-PerCP

IFN-PE

104

104

103

103

102

102

101

101100

100

CD4-PerCP

Il-17-FITC

104

104

103

103

102

102

101

101100

100

104103102101100

FVD-660

Side

Scatte

r

7000

9000

3.7% 56.0%

4.2% 36.1%

1.3% 32.5%

7.4% 58.8%

5000

3000

1000

0104103102101100


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21

FlowCellect Hman Memory B cell

Identication Kit

We have developed a multi-parameter ow cytometry

assay or monitoring human memory B cell unction. Merck

Millipores FlowCellect Human Memory B Cell Identifcation

Kit includes three directly conjugated antibodies: Anti-

HumanCD5-FITC,Anti-HumanCD19-APC,andAnti-

Human CD27-PE, along with optimized assay buers to

provide researchers the ability to phenotypically distinguish

cell types. All FlowCellect kits are optimized on guava

bench top ow cytometers. FlowCellect kits can be used on

any ow cytometer ollowing the same protocol providing

researchers a reliable and ully validated solution to study

and identiy human B cell unction right in the comort

o their own lab. All three antibodies provided in the kit

are careully titrated and optimized together to ensure

maximal perormance when run in multiplex, alleviating

the need or any additional optimization.

FEATuRED PRODuCT

Isolation and Identifcation o Human Memory B cells rom human PBMCs Human memory B cells are phenotypically identifed by

using specifc human CD markers: Human CD5, CD19, and CD27. CD27 has been identifed as the key marker or identiying memory

B cells. In (A), lymphocyte populations are gated, and B cells are shown using bivariate analysis plotting CD5+CD19+ (B). Memory B

cell subsets are also identifed by using a CD19+CD27+ (C).

In healthy patients, memory B cells represent approximately 30-60% o the B-cell pool. B-cell subpopulations are deective in

patients suering rom immuno-defciency disorders, showing a reduced number o circulating CD19+CD27+ memory B cells.


FlowCellect Hman Memory B Cell Identication KitAnti-CD5cloneDK23-FITC/Anti-CD19cloneHD37-APC/Anti-CD27cloneM-T271-PE

25 test FCIM025159

FlowCellect Human B Cell FAS KitCD19-FITC,CD45-PerCPofCocktail,Anti-HumanCD95(FAS)-PE/IsotypecontrolmouseIgG1-PE

100Tests FCCH100137


FlowCellect Mose Breg Identication KitAnti-HumanCD5,clone53-7.3-APC/Anti-MouseCD19,clone1D3-FITC/Anti-MouseCD1d,clone1B1-PE/Anti-MouseCD16/CD32,clone93Puried

25 Tests FCIM025154


FlowCellect Hman Lymphocyte ZAP-70 Characterization KitAnti-CD5-FITC/Anti-CD19-APC/Anti-ZAP-70-PE

25 Tests FCIM025122

Hman: B-cell

Mose: B-cell

Hman: T-cell Signaling

A.

Forward Scatter

Side

Sca

tter 7000

7000

9000

9000

Lymphocyte

5000

5000

3000

3000

1000

10000

0

B.

APC-CD19

FITC-CD5

CD5+B-cell

104

104

103

103

102

102

101

101100

100

C.

APC-CD19

PE-CD27

104

104

103

103

102

102

101

101100

100

Memory B-cell

B-cell

T-cell Signaling Kit


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22

Immne Cell Health (Apoptosis)

FlowCellect Hman T-Cell MitoDamage Kit

Merck Millipores FlowCellect T Human T cell MitoDamage

Kit includes (1) Antibody Cocktail containing CD3-PECy5,

CD4-PE and CD8-FITC antibodies (2) MitoSense Red

(1,1,3,3,3,3 -Hexamethylindodicarbocyanine iodide),

a uorescent cationic dye that accumulates in the

mitochondria and is responsive to mitochondrial potential

changes and (3) 1X Assay buer BA solution to perorm the

assays. The kit can thus distinguish multiple populations (1)

CD4THelpercellsand%ofthesecellswhichshowintact

mitochondrialmembranepotential(2)%ofCD4THelper

cells with dissipated mitochondrial membrane potential (3)

CD8cytotoxicTCellsandthe%ofthesecellsthatshow

intact mitochondrial membrane potential change and (4)

CD8CytotoxicTcellsand%ofthesecellsthatdemonstrate

dissipated membrane potential. The kit thus provides a

complete picture o T cell mitochondrial perturbation

status and its response or inducer treatment conditions

ordiseases.Theentireassaycanbeperformedin30min

a simple no wash manner without loss o apoptotic cells

when using PBMCs.

FEATuRED PRODuCT


FlowCellect Hman T Cell Apoptosis Kit(CD8-FITC,CD4-PE,CD3-PECy5ofcocktail/AnnexinV,CF647Reagent)

100Tests FCCH100138

FlowCellect Hman T Cell MitoDamage Kit(CD8-FITC, CD4-PE, CD3-PECy5 o cocktail/ MitoSense Red)

100Tests FCCH100139

FlowCellect Hman CD8 T Cell FAS Kit(CD8-FITC,CD3-PECY5ofCocktail/Anti-HumanCD95(FAS)-PE/IsotypecontrolmouseIgG1-PE)

100Tests FCCH100140

FlowCellect Hman T Cell Activation Kit(CD4-FITC,CD69-PE,CD3-PECY5,CD8-APCofcocktail) 100Tests FCCH100141

FlowCellect Hman CD4 T Cell FAS Kit(CD4-FITC,CD3-PECY5ofcocktail/Anti-HumanCD95(FAS)-PE/IsotypecontrolmouseIgG1-PE)

100Tests FCCH100154

FlowCellect Hman T Cell Caspase 8 Kit(CD4-PE, CD3-PECy5, CD8-APC o cocktail/ Caspase 8 FAM)

100Tests FCCH100155

FlowCellect Hman T Cell Caspase 9 Kit(CD4-PE,CD3-PECy5,CD8-APCofcocktail/Caspase9FAM)

100Tests FCCH100156

FlowCellect Hman T Cell Caspase 3/7 Kit(CD4-PE, CD3-PECy5, CD8-APC o cocktail/ Caspase 3/7 FAM)

100Tests FCCH100157

FlowCellect Hman B Cell FAS Kit(CD19-FITC,CD45-PerCPCocktail,Anti-HumanCD95(FAS)-PE/IsotypecontrolmouseIgG1-PE)

100Tests FCCH100137

Gava Cell Toxicity Kit(Guava CFSE/ CellToxicity 7-AAD)

100Tests 4500-0230

Hman

1 2

A

C

B

D

CD3-PECy5 CD4-PE

SSC

CD8

FITC

MitoSense Red

untreated Diamide

MitoSense Red

MitoSense Red

MitoSense Red

CD8+

FITC

CD4-PE

CD8+

FITC

CD4-PE

Mitochondrial membrane

depolarization in apoptotic

CD4 and CD8 T cells

PBMCs were treated overnightwith and without 50 M

diamide and analyzed with

the FlowCellect T Cell

MitoDamage Kit. Plots show

the percentage o positive

cells or CD3, CD4 and CD8

subpopulations (1, 2) and

the mitochondrial potential

status o CD4 and CD8 T cell

populations or untreated

(A, C) and treated samples

(B, D). Cells with intact

mitochondrial potential

exhibit higher Red2 or

MitoSense red uorescence

(as in untreated control),while cells with depolarized

mitochondria demonstrate

a downward shit in their

mitochondrial potential.


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23

Milli-MarkConjugated Antibodies

Advance your ow cytometry analysis with Merck

Millipores growing selection o directly conjugated

primary antibodies. As a part o our complete benchtop

ow cytometry solution including instruments, sotware,

service, and reagents, Milli-Mark uorescently-labeled

antibodies are specifcally designed, optimized and

validated or ow cytometry applications.

Stem Cell

Fixed and permeabilized2102Ep embryonal carcinoma

cells were stained with Milli-

Mark anti-hOct4-Alexa Fluor

488 (FCMAB113A4, green

histogram) or IgG-Alexa Fluor

488 (grey histogram) at 1:100

or 1 hour and then analyzed

by ow cytometry.

Description Reactivity Host Qty Catalog No.

Anti-hOCT4-Alexa Fluor 488 Human Mouse 100tests FCMAB113A4

Mouse

Anti-SSEA-4-PE Human Mouse 100tests FCMAB116P

Anti-Sox2-FITC Human Mouse 100tests FCMAB112F

Mouse

Rat

Anti-TRA160-FITC Human Mouse 100tests FCMAB115F

Anti-HESCA-1-FITC Human Mouse 100tests FCMAB111F

Mouse

Anti-SSEA-1-PE Human Mouse 100tests FCMAB117P

Mouse

RatAnti-SSEA3Alexa488,cloneMC-631 Human Rat 100tests FCMAB141A4

Anti-Human Nuclei -FITC, clone 3E1.3 Human Mouse 100tests FCMAB157F

Anti-BMP-7-FITC,clone2A10 Human Mouse 100tests FCMAB135F

Anti-mOCT4-AlexaFluor488,clone7F9.2 Human Mouse 100tests FCMAB124A4

Mouse

Anti-TRA-1-81-FITC,clone TRA-1-81 Human Mouse 100tests FCMAB132F

Anti-TRA-2-49-FITC,cloneTRA-2-49/6E Human Mouse 100tests FCMAB133F

Anti-SRF-FITC, clone 1E1 Human Mouse 100tests FCMAB137F


Anti-phospho-Bcl-2(Ser70)-PE,

clone69-10C-2-10C-18

Human Rabbit 100tests FCMAB140P

Rat-a-Caspase-2-FITC Human Rat 100tests FCMAB158F

Milli-Mark: Stem Cell

Milli-Mark: Apoptosis and Cancer

200

180

160

140

120

100

80

60

40

20

010e0 10e1 10e2 10e3 10e4

Count

hOCT4-Alexa Fluor 488


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Anti-MAD2A-FITC,clone17D10 Human Mouse 100tests FCMAB150F

Anti-Y14-FITC, clone 4C4 Human Mouse 100tests FCMAB151F

Anti-FXR1-FITC,clone6BG10 Human Mouse 100tests FCMAB152F

Anti-SMN-FITC, clone 2B1 Human Mouse 100tests FCMAB153F

Anti-CAF1p150-FITC,cloneSS1,1-3 Human Mouse 100tests FCMAB145FAnti-TBX21/T-Bet-FITC Human Mouse 100tests FCABS131F

Anti-RPA2p34-FITC,cloneRPA201-46 Human Mouse 100tests FCMAB143F

Anti-RPA1p70-FITC,cloneRPA9,1-30 Human Mouse 100tests FCMAB144F

Anti-BMI1-FITC, clone AF27 Human Mouse 100tests FCMAB149F

Anti-RBMS1-FITC, clone 4D11 Human Mouse 100tests FCMAB123F

Anti-c-Jun-FITC,clone6E4 Human Mouse 100tests FCMAB122F

Mouse

Rat

Anti-RBMS1-FITC, clone 4D11 Human Mouse 100tests FCMAB125F

Mouse

Anti-BrdU-Alexa 488 Human Mouse 100tests FCMAB101A4

Phospho-ATM(Ser1981)-PE Human Mouse 100tests FCMAB110PMouse

Rat

PhosphoH3(Ser10)-Alexa488 Human Mouse 100tests FCMAB104A4

Phospho-SMC1(Ser957)-Alexa488 Human Mouse 100tests FCMAB108A4

Bovine

Xenopus

Anti-Cyclin B1 - PE Human Mouse 100tests FCMAB102P

Mouse

Milli-Mark: Epigenetics and Gene Reglation

Epigenetics & Gene

Reglation

HeLa cells were stained with

either Milli-Mark anti-Y14-

FITC, clone 4C4 (FCMAB151F,

green histogram) or with

IgG2b isotype control antibody

(grey histogram) and analyzed

using ow cytometry.

200

180

160

140

120

100

80

60

40

20

0

10e0 10e1 10e2 10e3 10e4

Count

Green Fluorescence


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Anti-CD1a-FITC, clone NA1/34 Human Mouse 100tests FCMAB166F

Anti-CD2-FITC,cloneMT910 Human Mouse 100tests FCMAB167F

Anti-CD3-PECy5, clone UCHT1 Human Mouse 100tests FCMAB169C5

Anti-CD4-FITC,cloneMT310 Human Mouse 100tests FCMAB170F

Anti-CD8-FITC, clone DK25 Human Mouse 100tests FCMAB176F

Anti-CD11c-FITC,cloneKB90 Human Mouse 100tests FCMAB179F

Anti-CD13-FITC, clone WM-47 Human Mouse 100tests FCMAB180F

Anti-CD14-FITC, clone TUK4 Human Mouse 100tests FCMAB181F

Anti-CD16-FITC,cloneDJ130c Human Mouse 100tests FCMAB183F

Anti-CD19-APC,cloneHD37 Human Mouse 100tests FCMAB185AP

Anti-CD27-FITC, clone M-T271 Human Mouse 100tests FCMAB191F

Anti-CD45-PE,cloneF10-89.4 Human Mouse 100tests FCMAB118P

Anti-CD45RA-FITC,cloneMEM56 Human Mouse 100tests FCMAB126F

Mouse

Anti-CD56-PE,cloneMOC-1 Human Mouse Mouse 100tests FCMAB200P

Anti-CD57-FITC,cloneTB01 Human Mouse Mouse 100tests FCMAB201F


Anti-EGFR-PerCP, clone LA22 Human Mouse 100tests FCMAB129CP

Anti-mTOR-FITC, clone 2ID8.2 Human Mouse 100tests FCMAB154F

Anti-Ras-FITC,cloneRAS10 Human Mouse 100tests FCMAB148F

Anti-GbL/mLST8, clone 3E1.2, FITC Conjugate Human Mouse 100tests FCMAB121F

Mouse

Anti-Akt/PKB-AlexaFluor647,cloneSKB1 Human Mouse 100tests FCMAB128A6

Phospho-Erk1/2(Thr202/Tyr204,Thr185/Tyr187)-PE Human Rabbit 100tests FCMAB100P

Mouse

Rat

Anti-Ki-67-APC Human Mouse 100tests FCMAB103APPhospho-STAT1(Y701)Alexa488 Human Mouse 100tests FCMAB106A4

Mouse

Rat

Anti-b-catenin-PE, clone 7F7.2 Mouse Mouse 100tests FCMAB1209P

PhosphoSTAT5A/B(Y694/699)PEH Human Mouse 100tests FCMAB105P

Mouse

Bovine

Sheep

Milli-Mark: Infammation/Immnology

Milli-Mark: Cell Signaling


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26

Components o theGuava Flow Cytometry Solution

InstrmentsThe guava easyCyte ow cytometry systems are

uncomplicated instruments that deliver the power o

multiplexed cell analysisright on your benchtop. The

culmination o over a decade o ow cytometry expertise,

these instruments use minimal sample, generate less

waste, and are easier to use and maintain than traditional

ow cytometersall while providing the power you need

in the most compact ormat available. These advantages

are made possible by our patented microcapillary ow cell

technology.

Microcapillary Flow CytometryTechnologyAt the heart o every guava easyCyte system is a unique,

microcapillary ow cell that eliminates the need or sheath

uid. This translates into smaller samples, less reagents

and minimal wastesaving you both time and money.

Because the ow cell is sel-aligning and user-replaceable,

you can remove it yoursel at any time or cleaning and

maintenanceno more expense or downtime or service

visits. And by eliminating complicated microuidics,

weve created a compact to save valuable lab space and

eliminated much o the operational costs compared to

sheath-based instruments.

The patented Guava microcapillary allows or direct sampling,with no need or sheath uid and

complicated microuidics. The result is a compact, easy to use system whose operational costs are

a raction o a sheath-based instrument.

Sheath Fluid: None

Waste: < 80 mL per 8 hour run

Typical #Cells Per Protocol:

1,000 - 10,000 cells/test

Microcapillary

Sample

Laser

WasteLaser

Waste

Flow Cell

Sample

SampleFlow

Sheath Fluid: ~10 mL/test

Waste: 8,000 mL per 8 hour run

Typical #Cells Per Protocol: 100,000 -

1,000,000 cells/test

Traditional

Waste

SampleSheath Fluid

Laser

Laser

SheathFlow

InjectionTip

Waste

SampleFlow

Traditional sheath lid system

Gava-patented microcapillary system


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27

SPECIFICATIONS

Speciications

System easyCyte 5HT easyCyte 6HT easyCyte 6HT-2L easyCyte 8HT

Catalogue # 0500-4005 0500-4005 0500-4007 0500-4008

Option # N/A 0500-4006 N/A N/A

Laser Blue (488 nm) Blue (488 nm)

Blue (488 nm) and

Red(640nm)

Blue (488 nm) and

Red(640nm)

Laser Wattage 20mW 20mW 40mW 75 mW

FSC

SSC

Green 525/30nm 525/30nm 525/30nm 525/30nm

Yellow 583/26nm 583/26nm 583/26nm 583/26nm

Red1 680/30nm 680/30nm 690/50nm 690/50nm

NIR1 N/A 785/70nm N/A 785/70nm

Red2 N/A N/A 661/19nm 661/19nm

NIR2 N/A N/A N/A 785/70nm

Microcapillary Fluidics

Direct, Absolute Cell Counts

Automation96Well

and10Tubes

Mixing

Dell LatitudeE6520Laptop

with Intel Core

InCyte Sotware

guavaSuite Sotware Modules

Digital Signal Processing

gava easyCyte High Thoghpt Sampling Instrments

Small ootprint saves

valuable laboratory space

Width: 20.3 in (51.5 cm)

Depth: 23.4 in (59 cm)

Height: 10.0 in (25.4 cm)

(does not include laptop)

Wash vial oers a

high-pressure purge toeasily clear obstructions

rom the low cell

Waste vial collects less

than 80 mL o waste in a

typical 8-hour workday

Robotic sample tray

provides walk-away

automation or a 96-well

microplate and up to 10

sample tubes

Microcapillary low cell

requires no sheath luid andis user-replaceable

Up to six-color detection

made possible by one (blue) or

two excitation lasers (blue & red)


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28

SPECIFICATIONSSpeciications

System easyCyte 5 easyCyte 6 easyCyte 6-2L easyCyte 8

Catalogue # 0500-5005 0500-5005 0500-5007 0500-5008

Option # N/A 0500-5006 N/A N/A

Laser Blue (488 nm) Blue (488 nm)Blue (488 nm) and Red

(640nm)

Blue (488 nm) and Red

(640nm)

Laser Wattage 20mW 20mW 40mW 75 mWFSC

SSC

Green 525/30nm 525/30nm 525/30nm 525/30nm

Yellow 583/26nm 583/26nm 583/26nm 583/26nm

Red1 680/30nm 680/30nm 690/50nm 690/50nm

NIR1 N/A 785/70nm N/A 785/70nm

Red2 N/A N/A 661/19nm 661/19nm

NIR2 N/A N/A N/A 785/70nm

Microcapillary Fluidics

Direct, Absolute Cell Counts

Automation96Well

and10Tubes N/A N/A N/A N/A

Mixing N/A N/A N/A N/A

DellLatitudeE6520Laptop

with Intel Core

InCyte Sotware

guavaSuite Sotware Modules

Digital Signal Processing

gava easyCyte Single Sample Instrments

Small ootprint saves

valuable laboratory space

Width: 17.75 in (45.1 cm)

Depth: 17.25 in (44.5 cm)

Height: 8.75 in (22.2 cm)

(does not include laptop)

Wash vial oers a

high-pressure purge to

easily clear obstructions

rom the low cell

Single sample loader

Swivel arm unctional-

ity, holds two tubes and

allows instant acquisition

Waste vial collects less

than 80 mL o waste in a

typical 8-hour workday

Up to six-color detection

made possible by one (blue) or

two excitation lasers (blue

and red)

Microcapillary low cell

requires no sheath luid and is

user-replaceable


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29

SotwareYour specifc research needs are always changing and

Merck Millipores guava sotware is uniquely adaptable

to accommodate you at every level. The guavaSot

application-specifc modules have plug-and-play ormats

designed or our optimized reagents. For more exible,

user-defned ormats, InCyte sotware delivers many high-

level eatures which uniquely enable easy visualization

o data in a broad biological context. All our sotware

modules use the same intuitive user interace, to make it

easy to switch rom one ormat to another.

You can export data quickly to spreadsheets or any third-

party analysis ormat. Moreover, our sotware packages

can enable 21 CFR part 11 compliance.


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30

InCyte: Intitive

InCyte sotware brings a new level o analytical power to

ow cytometry. It is the frst solution designed to empower

every user to draw conclusions about the biological

signifcance o data.

Its intuitive, easy-to-use interace makes it possible to

visualize and compare up to eight data sets at once, withdrag and drop eatures to simpliy the set up o gating

strategies. Many high-level analysis eatures are already

built in. Entire experiments can now be analyzed and

viewed at once, in less time than it usually takes to analyze

a single sample. One o its uniquely powerul benefts is the

ability to display comparative results and the experiment

level, using eatures like heat maps and IC50

/EC50

curves

which allow easy target identifcation. Automated

compensation reduces the amount o time to analyze

complex multicolor assays by providing an automatic

correction o the spectral overlap o simultaneouslyanalyzed dyes. As a result, InCyte sotware can unction as

the primary data acquisition and analysis package or the

instrument.

D

E

FG H

Organize acquired data in

this panel

Easily create analysistemplates

Quicklylinktoandreview

previously analyzed data

Heat mapping allows rapid

visualizationofupto6

parameters at once, within a

single plate or across multiple

experiments

Construct heat maps or EC50

/

IC50

curves by selecting groups

o data and using slider bars to

set cut-os or threshold values

IC50

curves show inhibition

range o a dose-response

curve

Drag-and-drop gating

allows selection o speciic

populations or urther

analysis, with the ability to

highlight groups using colorback-gating

View up to 11 plots all at once,

with real-time plot adjustments


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Scepter 2.0The next generationo automated cell counting

Versatility in a Complete Package.MILLIPLEXmap magnetic bead-based immunoassay kits& MAGPIX instruments

PRODuCT HIGHLIGHTS

Scepter, the frst handheld automated cell counter,

brought portable and precise cell counting to the

palm o your hand

Scepter2.0nowexpandsyourabilitytoprecisely

count particles as small as 3 m in diameter or

compatibility with additional cell types such as: stem

cells, blood cells, and yeast.

Learn More:

www.millipore.com/scepter

Based on the Luminex xMAP technology and

Merck Millipores 25 years o experience, we provide

multiplex kits and ELISAs in key research areas,

enabling you to perorm multiple assays in a single

sample.

Learn more o our solutions or Cancer, Cellular

Metabolism, Neuroscience, and Toxicity at:

www.millipore.com/magbeads


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Merck Millipore and the M logo are trademarks o Merck KGaA, Darmstadt, Germany.

Cell Cycle Stop, ChemiScreen, easyCyte, InCyte, and Scepter are trademarks o Millipore Corporation.

guava ViaCount and Milli Mark are registered trademarks o Millipore Corporation

www.merckmillipore.com

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acebook.com/MerckMilliporeBioscience

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Innovative Solutions for Flow Cytometry Analysis Optimized Kits and Reagents PB3322ENEU

Documents

Transcript of Innovative Solutions for Flow Cytometry Analysis Optimized Kits and Reagents PB3322ENEU