MACS® Technology for the isolation and analysis of neural cells

Neuroscience

Isolate specific primary neural cellswithin just a few hours

Exclusive: myelin removal

Automated and fast dissociation of tissues

Molecular amplification and analysis from a single cell upwards

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Fig. 1: From stem cells to neurons and glia: neural lineage cells and their markers. A variety of cell surface antigens (highlighted in blue) can be magnetically labeled with MACS® MicroBeads and used to isolate neural stem and progenitor cells as well as populations from both neuronal and glial lineages.

Peripheral nervous system

Central nervous system

Neural stem cell CD133, Prominin-1

Neuronal-restricted precursor

PSA-NCAM

Glial-restricted precursor

A2B5

Neuron CD90 (Thy 1.2)

Oligodendrocyte Type-2 astrocyte

precursor

Type-1 astrocyte

Oligodendrocyte Type-2 astrocyte

Schwann cellCD271 (LNGFR)

MicrogliaCD11b

Neuroscience MACS® Technology for the isolation and analysis of neural cells

Contents

4 MACS® Neural Dissociation

Gentle and efficient dissociation of neural tissues

6 Myelin Removal Beads

Optimized sample quality for improved analysis

8 MACS® MicroBeads for neuroscience

Highly specific isolation of neural cell populations

12 Neuroimmunology

Isolation and analysis of leukocyte subsets

13 MACS® Cell Culture

Specialized and defined media for consistent results

14 Molecular analysis

Cutting-edge products and services for expression profiling of neural cells

16 References

Publications cited within this brochure

18 Order information

Place your order by fax, phone, or online!

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MACS® Technology has become the gold standard for cell separation since its introduction in 1989 and Miltenyi Biotec now offers more than 1,000 innovative products for biomedical and life science research.

MACS Technology allows the separation of magnetically labeled cells by the use of antibody-coupled magnetic MACS MicroBeads in combination with strong permanent magnets (MACS Separators) and specially designed MACS Columns.

The separation procedure takes less than one hour to complete and cells are ready for immediate use in downstream applications, such as flow cytometry or cell culture.

Target cells can be isolated by positive selection using cell-specific surface antigens or by negative selection through the depletion of unwanted cells. These two separation strategies can easily be combined to provide experimental flexibility during multiparameter isolation of cell subpopulations.

MACS MicroBeads• Highlyspecificmonoclonalantibodyconjugates

• Small(50nm),virus-sizednanoparticles

• Non-toxicandbiodegradable

• Colloidaltopreventsettlingandalloweasyhandlingwith short incubation times (15 minutes)

MACS MicroBeads for indirect magnetic labelingIndirect magnetic labeling with MACS MicroBeads allows any primary antibody to be used for cell separation. Monoclonal or polyclonal primary antibodies to be used can be either unconjugated, biotinylated, or fluorochrome-conjugated.

Anti-Immunoglobulin MicroBeads

Streptavidin MicroBeads

Anti-Biotin MicroBeads

Anti-Fluorochrome MicroBeads

Fig. 3: MACS® MicroBeads available for indirect magnetic labeling.

Magnetic labeling

Cells of interest are labeled with MACS® MicroBeads in a short incubation step.

Magnetic separation

Labeled and unlabeled cells are separated in a MACS Column placed within the magnetic field of a MACS Separator. The flow-through is collected as the non-magnetic, negative fraction.

Elution of the labeled cell fraction

The separation column is removed from the magnetic field and the retained cells are flushed out. Both the labeled and unlabeled fraction can be completely recovered and used for downstream applications.

Fig. 2: MACS® Cell Separation

MACS® Technology by Miltenyi BiotecIntegrated solutions: from cells to molecular analysis

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gentleMACS™ Dissociator The gentleMACS™ Dissociator is a benchtop instrument for the automated dissociation of tissues. The two types of unique gentleMACS Tubes used with the instrument enable time-saving dissociation or homogenization of tissues within a closed system. C Tubes are used with tissue in order to prepare viable single-cell suspensions for further cell separation, analysis, and culture. M Tubes are used for the homogenization of tissues or cells for molecular applications. The instrument offers optimized gentleMACS programs for a variety of specific applications.

• CTubesproducesingle-cellsuspensionsfromtissues

• MTubesproducehomogenizedcellandtissueextracts

• Time-savingautomatedtissuedissociation

• Standardizedprocedure

• Highlevelofusersafety

• Sterilesamplehandling

• gentleMACSprogramsavailableforneuraltissue dissociation — visit www.miltenyibiotec.com to download special protocols

MACS® Neural DissociationGentle and efficient dissociation of neural tissues

Fig. 1: gentleMACS™ Dissociator Table 1: Selection of Neural Tissue Dissociation Kit subtype in relation to antigen compatibility for subsequent MACS® Cell Separation.

Neural Tissue Dissociation KitsThe Neural Tissue Dissociation Kits, which can be used in combination with the gentleMACS™ Dissociator, have been developed for the gentle yet efficient generation of single-cell suspensions from neural tissues. The enzymatic dissociation procedure yields high numbers of viable cells in less than one hour with the dissociated cells immediately ready for in vitro or in vivo applications.

Antigen compatibilityAntigen Species Cell type

Neural Tissue Dissociation Kit (P)

Prominin-1 mouse Neural progenitor cells

A2B5 human, mouse, rat Glial-restricted precursors

O4 1 human, mouse, rat Immature oligodendro-cytes, type-2 astrocytes

CD11b human, mouse Microglia

CD81 1 mouse, rat Microglia, macrophages,endothelial cells, glia

CD31 1 mouse Endothelial cells

Neural Tissue Dissociation Kit (T)

Prominin-1 2 mouse Neural progenitor cells

PSA-NCAM human, mouse, rat Glial-restricted precursors oligodendrocyte progenitors

CD24 mouse Neuronal precursors, ependymal cells

A2B5 2 human, mouse, rat Glial-restricted precursors

CD11b human, mouse Microglia

CD271 (LNGFR)

human Schwann cells, motor neurons

CD105 mouse Endothelial cells

Ter-119 mouse Erythrocytes

1 Slight epitope sensitivity with the use of the papain-based kit; therefore, a further dilution of the enzyme mix is recommended.2 Slight epitope sensitivity with use of the trypsin-based kit; therefore, use of the papain-based kit recommended.

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MACS® Neural DissociationGentle and efficient dissociation of neural tissues

Single-cell suspensions from neural tissues in less than one hour

The dissociation kit is available in three formats: a papain-based Neural Tissue Dissociation Kit (P), a trypsin-based Neural Tissue Dissociation Kit (T), and the Neural Tissue Dissociation Kit – Postnatal Neurons that is optimised for dissociation of postnatal neural tissue.

• Two-stepenzymaticdissociationprocedure

• Gentletocellsforhighcellviability

• Suitableforembryonicandadultneuraltissue

• Suitableforwholebrainandtissuesections

• Availableinpapain,trypsin,orpostnatal-specificformat

• OptimizedforsubsequentMACSCellSeparation

2 min

15 min

Fig. 2: Dissociation of neural tissues using Neural Tissue Dissociation Kits.

10 min

20 min

10 min

Cut brain into small pieces, collect in HBSS and centrifuge.

Add pre-warmed Enzyme Mix 1 and incubate 15 min at 37 ° C.

Add Enzyme Mix 2 and mix gently

Dissociate mechanically by trituration with a wide-tipped glass pipette. Incubate 10 min at 37 ° C.

Repeat trituration with narrow-tipped glass pipettes until no tissue pieces remain.

Apply to cell strainer and wash twice with HBSS

(Optional) Myelin removal:

Process cells immediately, e.g. MACS® Cell Separation.

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Removal of myelin debris increases efficiency of MACS SeparationsMyelin is a specialized membrane that ensheaths and insulates axons in the peripheral and central nervous system. Large quantities of myelin debris are generated when neural tissue is dissociated. Magnetic cell isolation and immunostaining of dissociated cells are both considerably impaired by myelin particles.1,2

Myelin Removal Beads allow the specific removal of myelin debris from single cell suspensions during sample preparation leading to higher purity and increased recovery of target cells. Myelin removal is therefore recommended prior to cell sorting or antibody staining.

Myelin Removal BeadsOptimized sample quality for improved analysis

Fig. 1: Removal of myelin debris from single cell suspensions with Myelin Removal Beads Postnatal (P22) mouse brains were dissociated using the Neural Tissue Dissociation Kit (P) and the resulting single-cell suspensions analyzed by flow cytometry either before or after treatment with Myelin Removal Beads. Top panel, single-cell suspensions derived from mouse brain consist of large amounts of myelin membrane fragments and only 4% cells. Middle panel, particles outside the selected region stain with anti-O1, anti-O4 and anti-GalC antibodies that recognize glycolipids specifically contained within myelin membranes. Bottom panel, Myelin Removal Beads used with LD Columns and VarioMACS Separators efficiently remove myelin debris.

Before depletion

Forward scatter

Si

de

scat

ter

Forward scatter

Sid

e sc

atte

r

Anti-O1, -O4, -GalC staining

After Myelin Removal Beads

Forward scatter

Si

de

scat

ter

Forward scatter

Sid

e sc

atte

r

Forward scatter

Si

de

scat

ter

Forward scatter

Sid

e sc

atte

r

4.2%

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Fig. 2: Removal of myelin debris increases efficiency of MACS SeparationsPostnatal (P22) mouse brains were dissociated using the Neural Tissue Dissociation Kit (P) prior to MACS Separations using Anti-Prominin-1 MicroBeads. 1×107 cells from the single-cell suspension were then used

either directly for cell separation or were submitted to myelin depletion using Myelin Removal Beads prior to cell separation. Recovery of cells is higher in samples treated with Myelin Removal Beads prior to cell separation.

Original fraction

Forward scatter

Prom

inin

-APC

Negative fraction

Positive fraction with threshold

Forward scatter

Prom

inin

-APC

Forward scatter

Prom

inin

-APC

0.17%

0.04%

Forward scatter

Pr

om

inin

-AP

C

80.0%

Without myelin removal

Original fraction

Forward scatter

Prom

inin

-APC

Negative fraction

Positive fraction with threshold

Forward scatter

Prom

inin

-APC

Forward scatter

Prom

inin

-APC

With myelin removal

4.58%

0.43%

95.0%

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MACS® MicroBeads for neuroscienceHighly specific isolation of neural cell populations

CD133 — the human pluripotent stem cell markerCD133 is regarded as an important marker for the identification and isolation of primitive stem and progenitor cells from both hematopoietic and nonhematopoietic tissues.

CD133+ cells are also found in the human fetal brain and are regarded as neural stem and progenitor cells capable of forming self-renewing neurospheres in vitro and of differentiating into both neurons and glia.3,4,5

Furthermore, human CD133+ cells differentiate into fully integrated neurons and glial cells as well as astrocytes and endothelial cells when injected into mice.4,6,7

Mouse cerebellar neural stem cells express prominin-1Multipotent neural stem cells in the murine postnatal cerebellum can be isolated based on their expression of prominin-1, the murine ortholog of CD1338, in conjunction with their lack of expression of certain lineage-indicative markers (PSA–NCAM–, TAPA-1–, O4–).9 Purified prominin-1+/lineage– cells form self-renewing neurospheres in vitro and can differentiate into astrocytes, oligodendrocytes, and neurons. They can also regenerate each of these lineages after transplantation into the cerebellum.

Fig. 1: Isolation of CD133+ neural stem cellsCD133+ cells isolated from mobilized PBMCs were cultured for 3–5 weeks.Adherent cells differentiated into oligodendrocyte-like cells and stained for GFAP (FITC), MBP (Cy3), and nuclei (DAPI). (Courtesy of Dr. Kuçi, Tübingen, Germany)

Fig. 2: Prominin-1+ cells form neurospheres in culture Prominin-1+ cells were isolated from the cerebellum of postnatal day 1 mice using the Neural Tissue Dissociation Kit (P) followed by Anti-Prominin-1 MicroBeads.

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MACS® MicroBeads for neuroscienceHighly specific isolation of neural cell populations

PSA-NCAM is a marker of neuronal-committed progenitor cellsAnti-PSA-NCAM MicroBeads recognize polysialic acid (PSA) which is linked to the extracellular domain of the neural cell adhesion molecule (NCAM, CD56) in vertebrates.10

PSA-NCAM is predominantly expressed in embryonic and neonatal neural tissue.11 The expression of PSA-NCAM in mammalian adult brain is restricted mainly to areas that retain neurogenic potential, such as the subventricular zone (SVZ)12 and the dentate gyrus of the hippocampus.13 PSA-NCAM serves as a marker for immature neuronal-committed progenitor cells within these neurogenic areas.

Sequential sorting of neuronal precursors using MACS® TechnologyNeuronal-committed precursors can be isolated from the SVZ of the mammalian brain in a two-step procedure.14,15 Firstly, glial precursors can be depleted by their specific expression of A2B5 and the unlabeled cells collected in the flow-through can then be subjected to positive selection of neuronal precursors according to their expression of PSA-NCAM.

Isolated, highly purified PSA-NCAM+/A2B5– neuronal-committed precursors can be confirmed by staining with Beta III tubulin (TUJ1) and A2B5 antibodies as shown in Figure 3.

Fig. 3: Neuronal precursor isolation using MACS® Technology

TUJ1 A2B5

TUJ1 A2B5

Cells of the subventricular zone before separation

Magnetic labeling of A2B5 antigen upon surface of non-target cells

Magnetic separation and depletion of A2B5+ cells from flow-through fraction

Magnetic labeling of PSA-NCAM upon surface of target cells

Magnetic separation and retention of target cells within column

Elution of target cells from the column gives highly purified PSA-NCAM+/A2B5– neuronal-committed precursors

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A2B5 — a marker for glial-committed precursor cellsAnti-A2B5 MicroBeads recognize the c-series ganglioside antigen A2B5 and have been developed for the isolation of glial progenitor cells from embryonic or adult human, murine, or rat tissue by magnetic separation.15-22

Glycosphingolipids, including gangliosides, are considered to be useful cell-surface markers for monitoring neural stem and precursor cells as well as their progeny and have been utilized as specific markers to define cells of the oligodendrocyte lineage during development. A2B5 antigens are also expressed during development of glial precursor cells known as O-2A progenitor cells that differentiate into type-2 astrocytes and oligodendrocytes.23

CD11b — a marker for microglial cell isolationCD11b is widely used as a marker for microglial identification and is suitable to use for their immunomagnetic isolation from mouse brain tissue.24-27

Fig. 2: CD11b+ microglial cell isolation using CD11b (Microglia) MicroBeadsWhole-brain tissue from P1 mice was first dissociated to single cells using the Neural Tissue Dissociation Kit (T) and then microglia were isolated using CD11b (Microglia) MicroBeads. Cell samples were stained both before separation (top panel) and after separation (bottom panel) with CD11b-APC and CD45-FITC. A purity of approximately 91% was achieved for CD11b+CD45+ cells after magnetic separation.

CD11b-APC

CD

45-F

ITC

CD11b-APC

CD

45-F

ITC

Before separation

Positively selected CD11b+ microglia

Fig. 1: Cultured astrocytes derived from mouse brainCD1 mouse cortices (P1) were dissociated into a single cell suspension using the MACS Neural Tissue Dissociation Kit (P) in combination with the gentleMACS™ Dissociator. Target cell yield and purity were improved by the use of Myelin Removal Beads and the culture was maintained in MACS® Neuro Medium supplemented with MACS Supplement B27 PLUS. Astrocytes were labeled with an anti-GFAP antibody (FITC; green) and cell nuclei visualized by DAPI staining (blue).

MACS® MicroBeads for neuroscienceHighly specific isolation of neural cell populations

CD11b-APC

CD

45-F

ITC

CD11b-APC

CD

45-F

ITC

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Fig. 3: Isolation of Schwann cellsRat Schwann cells were isolated by MACS® Separation using CD271 (LNGFR) MicroBeads. Cells from both positive and negative fractions were then processed for immunofluoresence and stained with anti-CD271 (red) and Hoescht 33342 (blue).

CD271 (LNGFR) positive fraction following MACS® Separation

CD271 (LNGFR) negative fraction following MACS® Separation

Purification of Schwann cells using CD271 (LNGFR) as a selection markerSchwann cells can be successfully isolated from adult peripheral nerve biopsies according to their expression of CD271 (LNGFR, p75NTR).28,29 A Schwann cell purity of around 95% can be achieved using CD271 MicroBead Kits and this method only requires a couple of hours — a dramatic reduction in time when compared with standard culture methods.

Visit www.miltenyibiotec.com to download our protocol for the preparation and MACS Separation of Schwann cells.

CD90.2 for isolation of retinal ganglion cellsThe mouse CD90.2 antigen is expressed at high levels by retinal ganglion cells, thymocytes, and peripheral T cells. Mouse CD90.2 MicroBeads are suitable for the positive selection of mouse retinal ganglion cells from single-cell suspensions of retinal tissue and do not cross-react with Thy1.130-34. CD90.2 is expressed at lower levels on early hematopoietic stem cells in the bone marrow, interepithelial cells, and in skin.

MACS® MicroBeads for neuroscienceHighly specific isolation of neural cell populations

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Neuroinflammation and the neuroimmune response, such as the release of cytokines and proinflammatory chemokines by microglia and neural precursor cells, have been observed in a number of neurodegenerative diseases such as HIV-associated dementia35,36, multiple sclerosis37-39, Alzheimer’s disease40,41, and Parkinson’s disease.42

A broad range of cytokine products is available in addition to the comprehensive portfolio of cell separation reagents for the isolation of immune cells. A small number of products for neuroimmunology are provided as examples within the product tables — please find a more comprehensive list at:

www.miltenyibiotec.com

CytokinesA broad range of cytokines is available in premium grade format for high, well-defined activity as well as in research grade format for routine cell culture supplementation.

For a complete list visit: www.miltenyibiotec.com/cytokines

NeuroimmunologyIsolation and analysis of leukocyte subsets

Fig. 1: Example of a cell separation using the CD4+ T Cell Isolation Kit IIIsolation of untouched CD4+ T cells from PBMCs using the CD4+ T Cell Isolation Kit II and an LS Column. Cells were fluorescently stained with CD3-FITC and CD4-PE and analyzed by flow cytometry.

Before separation

Isolated CD4+ T cells

CD

3-FI

TC

CD4-PE

CD

3-FI

TC

CD4-PE

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MACS® Neuro MediumMACS® Neuro Medium has been optimized for the culture of neural cells following tissue dissociation with Neural Tissue Dissociation Kits and subsequent cell isolation by magnetic cell sorting.

MACS® Neuro Medium is a serum-free cell culture medium that promotes optimal growth and long-term survival of cells derived from the mouse, rat, and human central or peripheral nervous system.43,44

MACS® Neuro Medium is supplemented with growth factors suitable for the formation of neurospheres from magnetically isolated neural stem and progenitor cells. It is also suitable for the culture of magnetically isolated neuronal and glial precursors or differentiated cells derived from various tissues of the developing, postnatal and adult nervous system.

MACS® Supplement B27 PLUSMACS® Supplement B27 PLUS is a chemically-defined, serum-free supplement based on published formulations of B27 with re-optimized formulations of added components,45 and is recommended for low-density plating, long-term viability, and growth of CNS and PNS neurons. When added to MACS® Neuro Medium, MACS® Supplement B27 PLUS supports the growth of neuronal cells without an astrocyte feeder layer as well as the growth of neuronal tumor cell lines.

MACS® Cell CultureSpecialized and defined media for consistent results

Fig. 3: PSA-NCAM+ neuronal precursor cellsPSA-NCAM+ cells were stained with anti-GAD67 antibody after 7 days of culture in MACS Neuro Medium supplemented with MACS Supplement B27 PLUS.

Fig. 4: A2B5+ glial precursor cellsA2B5+ cells were stained with anti-GFAP antibody after 7 days of culture in MACS Neuro Medium supplemented with MACS Supplement B27 PLUS.

Fig. 2: Prominin-1+ neurospheresLight microscopic picture of neurosphere formation following magnetic cell sorting using Anti-Prominin-1 MicroBeads and 7 days of culture in MACS Neuro Medium supplemented with MACS Supplement B27 PLUS. Cells were prepared from CD1 mouse brain (P3) using the Neural Tissue Dissociation Kit (P).

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MACSmolecular provides a highly innovative range of products and services with a strong focus on microRNA and gene expression profiling. Sensitive downstream analyses are especially important when working with scarce and/or sensitive neural cell populations.

Easyprobe Transcription TemplatesIn situ hybridization and northern blot analysis are common and powerful methods for the detection and quantification of mRNA from different cells and tissues. It is essential to use single-stranded RNA probes that have a high sequence specificity to the target mRNA in order to achieve reliable and reproducible results. Often, the generation of these high-quality RNA probes turns out to be more tedious and time-consuming than expected.

EasyProbe Transcription Templates are ready-to-use, linear DNA templates that facilitate the generation of specific RNA probes for mouse neuronal-related mRNAs.

More detailed information on the EasyProbe Transcription Templates is available at: www.miltenyibiotec.com

Molecular analysisCutting-edge products and services for expression profiling of neural cells

DRAF

T

Fig. 2: In situ hybridization for cux-2 expression in mouse brain tissue Control hybridization with non-specific probe.

Fig. 1: In situ hybridization for cux-2 expression in mouse brain tissueHybridization with cux-2 probe generated by EasyProbe CUTL2, m Template.

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microRNA expression profiling — miRXplore™ Kits and ServicesExplore microRNA expression in human, mouse and rat neural cells with the miRXplore™ Microarray Kits and Services. Designed in collaboration with experts at the Rockefeller University46, the microarray covers more than 2,000 human, mouse, rat, and viral microRNA sequences and possesses rigorous internal control systems. Sequences differing by just one oligonucleotide can be reproducibly detected and re-ratios calculated with the use of the proprietary miRXplore Universal Reference.

DRAF

T

One-step mRNA isolation and in-column cDNA synthesis Premium mRNA can be isolated within 15 minutes directly from cells or tissues with the µMACS™ mRNA Isolation Kit. The µMACS One-step cDNA Kit combines efficient magnetic isolation of mRNA with revolutionary in-column cDNA synthesis. Purified cDNA can be generated from only a few to as many as 10⁷ cells.47

Agilent Whole Genome Microarray Services Certified serviceMiltenyi Biotec offers high-quality gene expression services based on Agilent whole-genome microarrays. The certified service team has fulfilled all criteria of Agilent’s microarray-based gene expression analysis certification program to ensure reliability of microarray results.

SuperAmp™ Service *The SuperAmp™ Service (available as an extension of the Agilent Microarray Services) can reliably amplify mRNA 1 million–fold from as little as one cell. The service is ideal for neural cells sorted with MACS® Technology, flow cytometry, or even from laser capture microdissected tissue.

* In combination with the Microarray Services only. The SuperAmp Service is not available for microRNA amplification.Fig. 3: a-Hyb™ Hybridization Station

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1. Pfenninger, C.V. et al. (2007) Cancer Res. 67: 5727–5736. 2. Tham., C.S. et al. (2003). Int. J. Dev. Neurosci. 21: 431–443. 3. Yu, S. et al. (2004) Biotech. Lett. 26: 1131–1136. 4. Belicchi, M. et al. (2004) J. Neurosci. Res. 77: 475–486. 5. Uchida, N. et al. (2000) Proc. Natl. Acad. Sci. USA 97: 14720–14725. 6. Tamaki, S. et al. (2002) J. Neuro. Res. 69: 976–986. 7. Cummings, B.J. et al. (2005) Proc. Natl. Acad. Sci. USA 102: 14069–14074. 8. Weigmann, A. et al. (1997) Proc. Natl. Acad. Sci. USA 94: 12425–12460. 9. Lee, A. et al. (2005) Nat. Neurosci. 8: 723–729. 10. Rougon, G. and Marshak, D.R. (1986) J. Biol. Chem. 261: 3396–3401. 11. Kiss, R.J. and Muller, D. (2001) Rev. Neurosci. 12: 297–310. 12. Doetsch, F. et al. (1997) J. Neurosci. 17: 5046–5061. 13. Seki, T. (2002) J. Neurosci. Res. 70: 327–334. 14. Seidenfaden, R. et al. (2005) Society for Neuroscience, Prog. No. 414.18. 15. Seidenfaden, R. et al. (2006) Mol. Cell. Neurosci. 32: 187–198. 16. Windrem, M.S. et al. (2004) Nat. Med. 10: 93–97. 17. Ruffini, F. et al. (2004) Am. J. Pathol. 165: 2167–2175. 18. Larsen, P.H. et al. (2006) J. Neurosci. 26: 2207–2214. 19. Larsen, P.H. et al. (2004) J. Neurosci. 24: 7597–7603. 20. Liu, A. et al. (2007) J. Neurosci. 27: 7339–7343. 21. Larsen, P.H. et al. (2003) J. Neurosci. 23: 11127–11135. 22. Strathman, F.G. et al. (2007) BMC Dev. Biology 7: 33. 23. Raff, M.C. et al. (1983) Nature 303 : 390–396. 24. Boddaert, J. et al. (2007) Am. J. Pathol. 170: 921–929. 25. Li, H. et al. (2006) J. Immunol. 177: 3644–3656. 26. Ndhlovu, L.C. et al. (2001) J. Immunol. 167: 2991–2999. 27. Schulter, D. et al. (2001) Brain Pathology 11: 44–55. 28. Vroemen, M. and Weidner, N. (2003) J. Neurosci. Methods 124: 135–143. 29. Manent, J. et al. (2003) J. Neurosci. Methods 123: 167–173. 30. Wu, D.Y. et al. (2003) Invest. Ophthalmol. Vis. Sci. 44: 2783–2790. 31. Shoge, K. et al. (1999) Neurosci. Lett. 259: 111–114. 32. Tsumamoto, Y. et al. (2002) Brain Res. 933: 118–129. 33. Mukai, S. et al. (2002) Jpn. J. Pharmacol. 89: 44–52. 34. Kerrison, J.B. et al. (2005) Mol. Vis. 11: 208–215. 35. Kelder, W. et al. (1998) Ann. Neurol. 44: 831–835. 36. Cinque, P. et al. (2005) J. Neuroimmunol. 168: 154–163. 37. Soerensen, T.L. et al. (2001) Eur. J. Neurol. 8: 665–672. 38. Mahad, D.J. et al. (2002) J. Neurol. Neurosurg. Psychiatry 72: 498–502.

ReferencesPublications cited within this brochure

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39. Scarpini, E. et al. (2002) J. Neurol. Sci. 195: 41–46. 40. Yamamoto, M. et al. (2005) Am. J. Pathol. 166: 1475–1485. 41. Xia, M.Q. et al. (2000) J. Neuroimmunol. 108: 227–235. 42. Nishimura, M. et al. (2003) Mov. Disord. 18: 953–955. 43. Brewer, G.J. et al. (1993) J. Neurosci. Res. 35: 567–576. 44. Brewer, G.J. (1997) J. Neurosci. Meth. 71:143–155. 45. Chen, Y. et al. (2008) J. Neurosci. Meth. 171: 239–247. 46. Landgraf, P. et al. (2007) Cell 129: 1401–1414. 47. Mack, E. et al. (2007) Cytometry A. 71: 404–409.

Find a comprehensive list of references citing the use of MACS Technology in neuroscience research available for download at:

www.miltenyibiotec.com/download/neuroreferences

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Order information Place your order by fax, phone, or online!

Product Specificity Order no.

MACS® MicroBeads for neural cell separation

CD133 MicroBead Kit human 130-050-801

Anti-Prominin-1 MicroBeads mouse 130-092-333

Anti-PSA-NCAM MicroBeads human, mouse, rat 130-092-966

Anti-A2B5 MicroBeads human, mouse, rat 130-093-388

CD90.2 MicroBeads mouse 130-049-101

CD11b (Microglia) MicroBeads human, mouse 130-093-634

CD271 MicroBead Kit (PE) human 130-092-819

CD271 MicroBead Kit (APC) human 130-092-283

Neural Tissue Dissociation Kit (P) +Anti-Prominin-1 MicroBeads

mouse 130-092-752

Examples of leukocyte cell separation reagents

CD3 MicroBeads human 130-050-101

CD4+ T Cell Isolation Kit II human 130-091-155

CD8+ T Cell Isolation Kit II human 130-091-154

CD14 MicroBeads human 130-050-201

Product Format available

MACS MicroBeads for indirect magnetic labeling

Anti-Fluorochrome MicroBeads FITC, PE, APC, Cy5/Alexa Fluor® 647, Cy 7

Anti-Fluorochrome MultiSort Kit* FITC, PE, APC

Anti-Biotin MicroBeads or MultiSort Kit* Biotinylated antibodies

Streptavidin MicroBeads Biotinylated antibodies

Anti-Immunoglobulin MicroBeads Monoclonal and polyclonal immunoglobulins

Product Specificity Format available

MACS Antibodies for neural cell identification and analysis

CD133 human PE, APC, Biotin, pure

Anti-Prominin-1 mouse PE, APC, Biotin, pure

Anti-PSA-NCAM human, mouse, rat PE, APC,

Anti-A2B5 human, mouse, rat PE, APC, Biotin, pure

CD90.2 mouse FITC, PE, APC

CD11b human, mouse FITC, PE, APC

CD271 (LNGFR) human PE, APC, Biotin

* MultiSort MicroBeads can be released from the cells for sequential cell sortings.

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Selected cytokines are available in a premium grade format for high, well-defined activity as well as in research grade quality.

For a complete list of available cytokines, please visit: www.miltenyibiotec.com/cytokines

For a complete list of MACS Media, please visit: www.miltenyibiotec.com/media

Product Order no.

microRNA expression profiling products

miRXplore Microarray Kit, 4 Microarrays 130-093-254

miRXplore Microarray Kit, 8 Microarrays 130-093-272

microRNA expression profiling services

miRXplore™ Universal Reference Service 160-001-161

miRXplore™ Microarray Service 160-001-143

miRXplore™ Additional Total RNA Extraction 160-001-162

mRNA & cDNA detection and isolation products

Easyprobe transcription templates Please inquire

µMACS™ mRNA Isolation Kit – Small Scale 130-075-201

µMACS™ mRNA Isolation Kit – Large Scale 130-075-101

µMACS™ One-step cDNA Kit 130-091-902

Agilent whole genome microarray services

Agilent Whole Genome Microarray Service, 4 × 44, One-color, various species

Please inquire

Agilent Whole Genome Microarray Service, 4 × 44, Two-color, various species

Please inquire

Custom Agilent Whole Genome One- and Two-color Microarray Services, various species

Please inquire

SuperAmp service 160-000-936

Product Order no.

MACS® Products for neural tissue dissociation & sample preparation

Neural Tissue Dissociation Kit (P) 130-092-628

Neural Tissue Dissociation Kit (T) 130-093-231

Neural Tissue Dissociation Kit – Postnatal Neurons NEW

130-094-802

gentleMACS™ Starting Kit 130-093-235

C Tubes, 25 tubes, single-packed 130-093-237

M Tubes, 25 tubes, single-packed

130-093-236

M Tubes, 50 tubes per bag 130-093-458

Myelin Removal Beads NEW 130-094-544

MACSmix™ Tube Rotator 130-090-753

FcR Blocking Reagent, human 130-059-901

FcR Blocking Reagent, mouse 130-092-575

Product Order no./ format

MACS Cell Culture Products

MACS Neuro Media NEW 130-093-570

MACS Supplement B27 Plus 130-093-566

Cytokines, human Activin A, BDNF, BMP-2, EGF, FGF-1, FGF-2 (basic FGF), FGF-4, IFN-γ, IGF-1, IGF-2, IL-1α, IL-1β, IL-2, IL-6, IL-8, NGF-β, TNF-α, VEGF

Cytokines, mouse EGF, G-CSF, GM-CSF, SCF, VEGF

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Unless otherwise specifically indicated, Miltenyi Biotec products and services are for research use only and not for diagnostic or therapeutic use. MACS is a registered trademark, and µMACS, a-Hyb, autoMACS, gentleMACS, MACSmix, MACSQuant, miRXplore, MultiMACS, PIQOR, PrepProtect, SuperAmp, and thermoMACS are trademarks of Miltenyi Biotec GmbH. Copyright © 2009 Miltenyi Biotec. All rights reserved.

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