LIFE SCIENCE PRODUCTS Catalogue - Euroclone group · PDF fileEMR069500...

LIFE SCIENCE PRODUCTS

Catalogue

4. Proteins Quantitation, Electrophoresis and Western Blotting

• • • •

Protein QuantitationAcrylamidesBuffers & ReagentsProtein Molecular Weight Markers Protein Gel StainingCustom IPG strips for 2D-electrophoresisVertical Gel Electrophoresis SystemsElectroblotting SystemsChemiluminescent Substrates for Western Blotting

• • • • •

Cap 4 Proteins....indd 1 4-10-2011 14:59:30

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Protein QuantitationEuroClone protein assay kits are formulations based on bicinchoninic acid (BCA) for the rapid and sensitive detection and quantitation of total protein content. The BCA method is faster and easier than Lowry, with much greater tolerance to interference from non-ionic detergents and Buffer salts.The BCA method combines the biuret reaction, i.e., the reduction of Cu2+ ions to Cu+ by proteins in an alkaline medium with complexation of the latter with bicinchoninic acid. The purple-colored Cu(BCA)2

3- complex displays a strong absorbance at 562 nm which is proportional to protein concentration over a broad working range (20-2.000 µg/ml for Quantum Protein and 0.5-20 µg/ml for Quantum Micro Protein).Protein concentrations are generally determined with reference to standards of a common protein such as Bovine Serum Albumin (BSA). If a more accurate quantitation of an unknown protein is required, the calibration curve is constructed using a protein similar to the unknown.

Quantum Protein & Quantum Micro Protein Assays

Features

Easy to use

Compatible with most common ionic and non-ionic detergents

Faster than Lowry method

Linear working range from 20-2.000 mg/ml (BCA) or 0.5-20 mg/ml (micro BCA)

Working solution extremely stable

Protocol flexibility to increase the sensitivity of the assay

Cuvette or microplate format

Protocol at a glance (Quantum Protein)

Prepare working solution (reagent A plus B ratio 50:1)

Add 2 ml working reagent to 100 ml sample

Incubate 30 min at 37 °C

Read at 562 nm

Cat. Description Format

EMP014500 Quantum Protein Kit (Linear working range 20-2000 µg/ml)

500 tube assays

EMP015480 Quantum Micro Protein Kit (Linear working range 0.5-20 µg/ml)

480 tube assays

Storage & Stability15 months at Room TemperatureShippingRoom Temperature


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Acrylamides

EuroClone offers ready-to-use acrylamide stock solutions for PAGE. For most applications in proteins electrophoresis, polyacrylamide gels are prepared from 30% or 40% stock solutions with a ratio Acrylamide/Bisacrylamide of 29:1 or 37.5:1.


EMR069250 Acrylamide/Bisacrylamide 29:1 Sol 30% MBG 250 ml

EMR069500 Acrylamide/Bisacrylamide 29:1 Sol 30% MBG 500 ml

EMR069001 Acrylamide/Bisacrylamide 29:1 Sol 30% MBG 1 L

EMR076250 Acrylamide/Bisacrylamide 37.5:1 Sol 30% MBG 250 ml

EMR076500 Acrylamide/Bisacrylamide 37.5:1 Sol 30% MBG 500 ml

EMR076001 Acrylamide/Bisacrylamide 37.5:1 Sol 30% MBG 1 L

EMR014500 Acrylamide/Bisacrylamide 37.5:1 Sol.40% MBG 500 ml

EMR014001 Acrylamide/Bisacrylamide 37.5:1 Sol.40% MBG 1 L

MBG= Molecular Biology grade

Buffers and Reagents

EuroClone can offer a complete choice of reagents and Buffers for either electrophoresis and Western Blot.The table below shows a selection of these reagents.


EMR084100 Albumin acetylated 100 mg

EMR085025V Albumin Fraction V - pH 5.2 25 g




EMR086001 Albumin Fraction V - pH 7.0 1 Kg

EMR086025 Albumin Fraction V - pH 7.0 25 g





EMR025100 Ammonium persulfate 100 g

EMR025250 Ammonium persulfate 250 g

EMR142000 Gelatin, powdered 5 kg

EMR142001 Gelatin, powdered 1 kg

EMR180001 Milk, non-fat dried 1 kg

EMR180500 Milk, non-fat dried 500 g

EMR194100 Ponceau S - 0,1% solution 100 ml

EMR194500 Ponceau S - 0.1% solution 500 ml

EMR228100 TEMED 100 ml



EMR242001 TTBS (Tween/Tris-Buffered salt solution) 1 L

EMR242250 TTBS (Tween/Tris-Buffered salt solution) 250 ml

EMR242500 TTBS (Tween/Tris-Buffered salt solution) 500 ml

EMR236001 Tris-Glycine Buffer, 10X 1 L

EMR236500 Tris-Glycine Buffer, 10X 500 ml

For the complete list of EuroClone Biochemicals please

refer to Chapter 8 or visit www.euroclone.it


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EMR244025 Tween 20 - 10% solution peroxide-free 25 ml




EMP011005 LiteAblot® PLUS 5000 cm2 membrane

EMP013001 LiteAblot® EXTEND 1000 cm2 membrane

EMP012001 LiteAblot® TURBO 1000 cm2 membrane

EPS010001 GelMateGhost 1 L sufficient for 40 mini gels

EPS011001 GelMateBLUE 1 L sufficient for 30 mini gels

For more details about protein gel stains and chemiluminescent susbtrates please see page

46/47, 57/59 or visit www.euroclone.it


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Protein Molecular Weight Markers

EuroClone Protein SHARPMASS™ Low, Broad and High are unstained ready-to-use protein molecular weight markers for precise molecular weight determination.Prestained SHARPMASS™ III and V are ideal for monitoring migration during SDS-PAGE and transfer efficiency in Western Blot procedures.

Protein SHARPMASS™ High (45-179 kDa)

Protein SHARPMASS™ High is a mixture of 5 purified proteins. The mixture is formulated to yield well-defined bands which after SDS-PAGE (run according to Laemmli) and staining with Coomassie Blue R-250 are approximately equal in color intensity.

Protein SHARPMASS™ Low (14.4-97 kDa)

Protein SHARPMASS™ Low is a mixture of 6 purified proteins. The mixture is formulated to yield well-defined bands which after SDS-PAGE (run according to Laemmli) and staining with Coomassie Blue R-250 are approximately equal in color intensity.

Protein SHARPMASS™ Broad (6.5-179 kDa)

Protein SHARPMASS™ Broad is a mixture of 9 purified proteins. The mixture is formulated to yield well-defined bands which after SDS-PAGE (run according to Laemmli) and staining with Coomassie Blue R-250 are approximately equal in color intensity.


EPS012500 Protein SHARPMASS™ High 2 x 250 μl

EPS013500 Protein SHARPMASS™ Low 2 x 250 μl

EPS014500 Protein SHARPMASS™ Broad 2 x 250 μl

Storage and StabilityOne year at –20 °C

ShippingBlue Ice


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Prestained Protein SHARPMASS™ III (19-117 kDa)

Prestained Protein SHARPMASS™ III is a mixture of 6 highly purified proteins covalently coupled to a blue chromophore. The marker is supplied ‘ready-to-use’ in loading Buffer.

Prestained Protein SHARPMASS™ V (11-250 kDa)

Prestained Protein SHARPMASS™ V is a mixture of 9 coloured proteins. These proteins are recombinantly produced in E.coli, highly purified and supplied ‘ready-to-use’ in loading Buffer. The protein concentrations are optimised to yield well-defined bands directly visible in SDS-polyacrylamide gels.Seven of the 9 proteins ranging in apparent molecular weight from approximately 11 kDa to 250 kDa are coupled with a blue chromophore and resolve into clearly defined, sharp bands when analysed by SDS-PAGE. A 72 kDa and 28 kDa protein is coupled with an orange dye and is serving as a reference band.


EPS022500 Prestained Protein SHARPMASS™ III 2 x 250 μl

EPS023500 Prestained Protein SHARPMASS™ V 2 x 250 μl

Storage and StabilitySix months at –20 °CShippingBlue Ice


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Protein Gel Staining

GelMateBLUE Permament backgroundless stain for protein gel

GelMateBLUE is a permanent staining ideal for any kind of protein gel. It allows the detection of as little as 10 ng of proteins. It contains the Coomassie G-250 dye, whose colloidal properties are responsible for the protein staining on polyacrylamide gels. GelMateBLUE stains only proteins, and bands can be seen directly during the staining process. After the staining step, a rinse in distilled water further enhances staining sensitivity and guarantees a clear background. The entire GelMateBLUE staining process can be completed without using any organic solvent to eliminate the excess of dye from the gel.

Features

Ready-to-use

Coomassie G-250 Dye

More sensitive than classic Coomassie Stain

No destaining step necessary

Mass Spectrometry compatible

Protocol at a glance

Fixing in 40% methanol and 10% acetic acid (1 hour)

Rinsing in distilled water

Staining with GelMateBlue (1 hour)


EPS011001 GelMateBLUE 1 L sufficient for 30 mini gels

Storage & StabilityOne year at Room TemperatureShippingRoom Temperature

GelMateBLUE is more sensitive than competitor colloidal Coomassie Stain.10 and 20 mg of total protein were loaded on 12.5 % SDS-PAGE. The gel was then stained with GelMateBLUE or with a competitor colloidal Coomassie stain.


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GelMateGhost Reversible negative stain for SDS protein gelEuroClone GelMateGhost provides rapid detection of proteins on mono-dimensional and bi-dimensional electrophoretic gels. GelMateGhost stains the empty matrix and leaves it clear where proteic bands remain. The result is a negative staining where proteins appear as clear bands in a milky background. The bands are visualized by placing the gel against a white light transilluminator.GelMateGhost is optimised for verifying the amount of proteins after electrophoresis before proceeding with further analysis or any kind of gel staining (Coomassie Blue, Silver Staining, Fluorescent Staining). It uses an easy, two-step procedure that requires very little time to complete. The white background is then removed with a simple rinse in distilled water and the gel can be re-stained by any permanent stain compatible with mass analysis. Alternatively, the gel can be destained with a Tris-glycine Buffer, allowing proteins to be eluted or transferred for Western Blotting. The eluted protein is free of dye contamination, making it ideal for downstream applications such as antibody generation or immunological detection. All reagents are provided ready to use, making this an ideal stain for multiple applications.

Features

Ready-to-use

Rapid Protocol

Fixing step not required

Compatible with a wide range of downstream applications

Mass Spectrometry compatible

Protocol at a glance

Gel rinsing in distilled water (5 min)

Staining in GelMateGost (1 hour)

Washing in distilled water


EPS010001 GelMateGhost 1 L sufficient for 40 mini gels

Storage & StabilityOne year at Room TemperatureShippingRoom Temperature


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CUSTOM IPG strips for 2D-electrophoresis

High reproducibility and better resolution!

• Choose narrow (pH 4-5.5) or medium (pH 4-7, pH 4-8) pH gradient, but if you need broad or particular ranges, ask for your personalized gradient. It can be available for you in a couple of weeks.• Increase loading capacity over two milligram for preparative and visualize more than 800 proteins in a single pH unit.• Ensure highly efficient sample entry and no more problems for small quantity of your sample.

Focus on inflammation proteins on serum: Coomassie staining, 2 milligrams in a 1.5 unit of pH.

Zoom on acidic proteins on bacterial extract: Coomassie staining 200 micrograms of bacterial extract.

Bacterial extract: Silver staining, 18 cm length, 50 μg.

Please find herebelow some of our custom IPG Strips Cat. Description Format

EMP050040 Custom IPG Strips pH 4-7 40 strips

EMP051040 Custom IPG Strips pH 4-5.5 40 strips

EMP052040 Custom IPG Strips pH 4-8 40 strips


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Vertical Gel Electrophoresis Systems

EuroClone vertical electrophoresis systems come in four different formats to accomodate quick or high-resolution separation for a wide range of sample numbers.

System Components• casting base with silicon mats• 4 glass plate pair sets• 2 or 4 Combs and spacer sets (depending on model)• 1 blind plate for single-gel runs• 2 replacement gaskets

Features

3-cathode technology and platinum-plated electrodes for exceptional electrical field uniformity and distortion-free gel runs

Centrally located flow cooling system made with alumina ceramic ensures optimal heat removal and outstanding separation efficiency even at high currents

Asymmetrical configuration of electrical contacts prevents mismatching of electrodes

Easy-to-use casting system: gel casting directly in the chamber assembly on casting base with silicone sealing mats for agarose-free sealing

“Blocking plate” for single-gel run


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Double Twin S

Features

Gel Size 10 x 10 cm

Footprint 17 x 14 x 13 cm

Sample Flexibility max. 24

Buffer Volume 250 ml

Cat. Description

EM45-1010 Double Twin S

System Components• Complete System with Internal Cooling Core• SuperSafe Cover with Attached Leads• 4 Glass Plates, 2 Notched Glass Plates• 2 Notched Ceram Plates• 2 Combs: 0.8 mm, 10 teeth• 2 Spacer Sets: 0.8 mm• 2 Replacement Gaskets• 1 Blocking Plate• 1 SpacerPlacer• 1 Casting Base

Accessories:Cat. Description

EM45-1010-10G Blank glass, 10 cm W x 10 cm H x 3/32” thick

EM45-1010-10RA Notched alumina, 10 cm W x 10 cm H x 1mm thick

EM45-1010-10R Notched glass, 10 cm W x 10 cm H x 3/32” thick

EM45-1010-10C Comb, 0.8 mm thick, 10 tooth, Sample volume: 50 µl

EM45-1010-10D Comb, 1.5 mm thick, 10 tooth, Sample volume: 90 µl







EM45-1010-CL Replacement Clamps, Set of 2

EM45-1010-SC Spacer Set, 0.8 mm thick

EM45-1010-SD Spacer Set, 1.5 mm thick

EM45-1010-CB Gel Casting Base Only

EM45-1010-CU Gel Casting Stand Includes Casting Base and Double Sided Precision-Machined Acrylic Block fitted with gaskets, positioning pins and clamps. Allows for casting of 10 cm W gels independently.

EM45-1010-CG Replacement gasket for Gel Casting Base

EM45-1010-GK Replacement gasket for UBC, set of 2


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Double Twin M

Features

Gel Size 16 x 14 cm




Cat. Description

EM45-1614 Double Twin M

System Components• Complete System with Internal Cooling Core• SuperSafe Cover with attached leads• 4 Glass Plates, 4 Notched Glass Plates• 4 Combs, 1.5 mm (2 x 10, 2 x 15 teeth)• 4 Spacer Sets, 1.5 mm• 2 Replacement gaskets• 1 Blocking plate• 1 Casting base

AccessoriesCat. Description

EM45-1614-14G Plain Glass Plate, 16 x 14 x 0.32 cm

EM45-1614-14R Notched Glass Plate, 16 x 14 x 0.32 cm

EM45-1614-10C 10 tooth Comb, 0.8 mm thick, Sample volume: 125 µl

EM45-1614-10D 10 tooth Comb, 1.5 mm thick, Sample volume: 234 µl







EM45-1614-PD Prep Comb, 2 tooth, 1.5 mm thick

EM45-1614-SC Spacer Set, 2 side spacers, 0.8 mm thick

EM45-1614-SD Spacer Set, 2 side spacers, 1.5 mm thick


EM45-1614-CB Casting Base Only

EM45-1614-CU Gel Casting Stand includes Casting Base and Double Sided Precision-Machined Acrylic Block fitted with gaskets, positioning pins and clamps. Allows for casting of 16 cm W gels independently.

EM45-1614-CG Replacement Gasket Set for 85-1614-GCB

EM45-1614-GK Replacement Gasket Set (2) for 85-1614 Upper Buffer Chamber


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Double Twin L

Features

Gel Size 20 x 20 cm

Footprint 30 x 24 x 17



Cat. Description

EM45-2020 Double Twin L

System Components• Complete System with Internal Cooling Core• SuperSafe Cover with attached leads• 4 Glass Plates, 4 Notched Glass Plates• 4 Combs, 1.5 mm (2 x 15, 2 x 20 teeth)• 4 Spacer Sets, 1.5 mm• 2 Replacement gaskets• 1 Blocking plate• 1 Casting base


EM45-2020-20G Blank Glass Plate, 20 cm x 20 cm x 0.32 cm

EM45-2020-20R Notched Glass Plate, 20 cm x 20 cm x 0.32 cm

EM45-2020-10C 10 tooth Comb, 0.8 mm thick

EM45-2020-10D 10 tooth Comb, 1.5 mm thick






EM45-2020-PD Prep Comb, 2 tooth, 1.5 mm thick

EM45-2020-CB Gel Casting Base Only

EM45-2020-CU Gel Casting Stand includes Casting Base and Double Sided Precision-Machined Acrylic Block fitted with gaskets, positioning pins and†clamps. Allows for casting of 20 cm W gels independently.

EM45-2020-CG Replacement Gasket Set (2) for Casting Base

EM45-2020-GK Replacement Gasket Set, Upper Buffer chamber, Pk 2

EM45-2020-SC Spacer Set, 2 side spacers, 0.8 mm thick

EM45-2020-SD Spacer Set, 2 side spacers, 1.5 mm thick


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Double Twin ExWs

Features

Gel Size 20 x 10 cm




Cat. Description

EM45-2010 Double Twin ExWs

System Components• Complete System with Internal Cooling Core• SuperSafe Cover with attached leads• 4 Glass Plates, 4 Notched Glass Plates• 4 Combs: 0.8 mm thick (2 x 15, 2 x 20 teeth)• 4 Spacer Sets: 0.8 mm• 2 Replacement gaskets• 1 Blocking plate• 1 Casting base


EM45-2010-10G Blank Glass Plate, 20 cm W x 10 cm H x 0.24 cm

EM45-2010-10R Notched Glass Plate, 20 cm W x 10 cm H x 0.24 cm










EM45-2010-CB Gel Casting Base†

EM45-2010-CU Gel Casting Stand includes Casting Base and Double Sided Precision-Machined Acrylic Block fitted with gaskets, positioning pins and clamps. Allows for casting of 20 cm W gels independently.

EM45-2010-CG Replacement Gasket Set, Casting Base

EM45-2010-GK Replacement Gasket Set, Upper Buffer chamber, Pk 2

EM45-2010-SC Spacer Set, 2 Side Spacers, 0.8 mm thick

EM45-2010-SD Spacer Set, 2 Side Spacers, 1.5 mm thick


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Electroblotting Systems

EuroClone Electroblotting Systems are designed for the transfer of proteins in conjunction with semi-dry or wetblot techniques. Electroblotting Systems of either types are available in two different sizes.

Wet Blotting Systems

EuroClone Wet Blotting Systems are designed for the effective and rapid transfer of molecules onto membranes.The platinum-plated titanium anode and stainless steel cathode provide an uniform electrical field for reproducible transfer results. A water circulation system with alumina ceramic wall located at the chamber base manages heat build up ensuring dependable results.


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Insight Mini Tank Electroblotter

Features

Transfer area 8.5 x 9.5 cm

Footprint 18.5 x 15.5 x 14.5 cm

Buffer volume 1300 ml

Blotting capacity 1 - 4 gels

Cat. Description

EM52-WEB-10 Insight Mini Tank Electroblotter

System Components:• Buffer chamber with durable plate electrodes• Platinum-plated Titanium anode• Stainless steel cathode• Lid with attached cables• 4 blotting cassettes• 8 fibre pads


EM52-WEB-SF Fibre Pads for Web-S, 10 x 12.5 cm, 4 pieces

EM52-WEB-SC Blotting Cassette for Web-S, incl. 2 fibre pads

Insight Large Tank Electroblotter

Features

Transfer area 18 x 20 cm


Buffer volume 4500 ml

Blotting capacity 1 - 2 gels

Cat. Description

EM52-WEB-20 Insight Large Tank Electroblotter

System Components:• Buffer chamber with durable plate electrodes• Platinum-plated Titanium anode• Stainless steel cathode• Lid with attached cables• 2 blotting cassettes• 4 fibre pads


EM52-WEB-MF Fibre Pads for Web-M, 18 x 24.5 cm, 4 pieces

EM52-WEB-MC Blotting Cassette for Web-M, incl. 2 fibre pads


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Semi-Dry Electroblotting Systems

EuroClone ’Semi-Dry’ Electroblotting Systems are designed for rapid and Buffer saving transfers of proteins and nucleic acids to membranes. The platinum-coated titanium anode and the high-grade stainless steel cathode form a uniform electrical field and ensure maximum transfer efficiencies. The asymmetrical configuration of electrical contacts prevents mismatching of electrodes.

Insight Semi-Dry Electroblotter Mini

Cat. Description

EM52-1010 Insight Semi-Dry Electroblotter Mini

System Components:• Transfer Area: 10 x 10 cm• Base with Platinium/Titanium Anode• Lid with Stainless Steel Cathode• Interlocking Power Cords• 3 Heavy Duty Knobs

Insight Semi-Dry Electroblotter Maxi

Cat. Description

EM52-2020 Insight Semi-Dry Electroblotter Maxi

System Components:• Transfer Area: 20 x 20 cm• Base with Platinium/Titanium Anode• Lid with Stainless Steel Cathode• Interlocking Power Cords• 3 Heavy Duty Knobs


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Chemiluminescent Substrates for Western Blotting

EuroClone can offer a complete range of chemiluminescent substrates to satisfy any need in terms of reliability, sensitivity and signal duration. In addition, all chemicals contained in these substrates have been carefully selected for safety: none of the components has been reported to be hazardous to human health.

LiteAblot® PLUS Enhanced Chemiluminescent Substrate

LiteAblot® PLUS is an extremely cheap chemiluminescent substrate more sensitive then classic ECL. The signal develops rapidly and it is stable for some hours. It is suggested for routinary Western Blot and it is ideal for the detection of standard protein expression levels. The substrate is easly prepared by mixing its two components, a luminol/enhancer solution and a peroxide solution, in a one-to-one ratio. This working solution is stable for up to 24 hours at RT.

LiteAblot® Plus is more sensitive than Competitor ECL Substrate.Recombinant human TNFa was serially diluted (40-5 ng) and electrophoresis was performed. The gels were transferred to nitrocellulose membranes, blocked and incubated with 1/1000 dilution of rabbit anti human TNFa antibody. After washing the membranes were incubated with 1/2000 dilutions of HRP-conjugated anti rabbit antibody. The membranes were washed again and then incubated with the substrate that was prepared according to the manufacturers’instructions. Blots were exposed for 1 and 5 min.

Features

Easy to use

High sensitivity

Stable signal


EMP011005 LiteAblot® PLUS 5000 cm2 membrane

Storage & StabilityOne year at 4 °CShippingRoom Temperature


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LiteAblot® EXTEND Long Lasting Chemiluminescent Substrate

LiteAblot® EXTEND has been specifically formulated to provide an intense and extremely stable chemiluminescent signal. For this reason this product is paricularly suggested to customers using cooled charge coupled device (CCD) technology. Considering the high sensitivity of this substrate compared to standard “ECL-like” products, it can be necessary to dilute primary and secondary antibodies much further.

Features

Excellent sensitivity

Long lasting signal (up to 24 hrs)


EMP013001 LiteAblot® EXTEND 1000 cm2 membrane


Images of a 50 pg dot blot incubated 5 minutes with LiteAblot® EXTEND.The signal has been captured with a NighOwl Luminograph (Berthold T-echnologies). Readings were taken at the indicated times after incubation, with an exposure time of 10 minutes, except for the last reading (1 hour).


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LiteAblot® TURBO Extra Sensitive Chemiluminescent Substrate

LiteAblot® TURBO is EuroClone chemiluminescent substrate with the highest sensitivity allowing the detection of very low amounts of proteins. Due to the extreme sensitivity of the substrate it is critical to optimize the primary and secondary antibody dilutions and the exposure times.

Features

Maximal sensitivity

Detection of low abundant poteins

Stable signal


EMP012001 LiteAblot® TURBO 1000 cm2 membrane



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StripAblot Stripping Buffer

StripAblot is a ready-to-use Buffer for the efficient stripping of Western Blot membranes. After a rapid incubation the stripped membrane can be re-probed with a new primary and secondary antibody and detceted with chemiluminescent substrates.

Features

Ready-to-use

Rapid protocol (5-15 minutes)

Mercaptoethanol free


EMP100500 StripABlot 500 ml

Storage & Stability15 months at RT or 4°CShippingRoom Temperature


Distributed by:

EuroClone S.p.A.Via Figino, 20/22 - 20016 Pero (MI) Italy +39 02 38195.1 - +39 02 38101465 [email protected] - www.euroclone.it EuroClone S.p.A. has a Quality System certified in compliance withUNI EN ISO 9001:2008 and NF EN ISO 13485:2004

EuroClone S.p.A. reserves the right to change product specification without prior notice

LIFE SCIENCE PRODUCTS Catalogue - Euroclone group · PDF fileEMR069500...

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