Revision 1

#38289Store at -20C

1 Kit

(8 x 20 microliters)

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

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3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
Olig2 (E6G6Q) XP® Rabbit mAb 65915 20 µl 35 kDa Rabbit IgG
Myelin Basic Protein (D8X4Q) XP® Rabbit mAb 78896 20 µl 12-18 kDa Rabbit IgG
CNPase (D83E10) XP® Rabbit mAb 5664 20 µl 47 kDa Rabbit IgG
PLP1 (E9V1N) Rabbit mAb 28702 20 µl 20-30 kDa Rabbit IgG
MAG (D4G3) XP® Rabbit mAb 9043 20 µl 100 kDa Rabbit IgG
MOG (E5K6T) XP® Rabbit mAb 96457 20 µl 46, 35, 28, 23 kDa Rabbit IgG
NG2/CSPG4 (E3B3G) XP® Rabbit mAb 43916 20 µl 250, 450 kDa Rabbit IgG
Sox10 (E6B6I) XP® Rabbit mAb 69661 20 µl 62 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The Oligodendrocyte Marker Antibody Sampler Kit provides an economical means of detecting proteins identified as oligodendrocyte markers by immunofluorescence and western blot. This kit includes enough primary antibodies to perform at least twenty IF tests or two western blot experiments per primary antibody.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/mL BSA, 50% glycerol, and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibodies.

Background

Oligodendrocytes are the myelinating glial cells of the central nervous system (CNS) (1). Myelin basic protein (MBP) is an abundant CNS myelin membrane protein that plays an important role in nerve myelination. MBP helps to adhere the cytoplasmic leaflets of adjacent oligodendrocyte membranes to one another (2). CNPase (2', 3'-cyclic nucleotide 3'-phosphodiesterase) is an enzyme highly expressed in oligodendrocytes and accounts for roughly 4% of the total myelin protein in the CNS (3). CNPase binds to tubulin heterodimers and plays a role in tubulin polymerization, and oligodendrocyte process outgrowth (4). Myelin proteolipid protein (PLP1) corresponds to the majority of myelin proteins in the CNS, providing support to axons and modulating axonal growth (5). Myelin-associated glycoprotein (MAG) is localized in oligodendroglial membranes of myelin sheaths where it plays a role in interaction between axons and glia, and has been shown to promote axonal protective effects (6,7). Chondroitin sulfate proteoglycan 4 (CSP4, NG2) is a type I membrane glycoprotein expressed by oligodendroglial precursor cells (OPCs) that plays a role in cell attachment and migration (8). SRY-box 10 (Sox10) is a high-mobility group transcription factor expressed throughout oligodendrocyte development required for myelin gene expression (9,10). Myelin-oligodendrocyte glycoprotein (MOG) is a type I membrane bound glycoprotein of the immunoglobulin superfamily that is enriched in the outer lamella of the myelin sheath (11). Olig2 is a basic helix–loop–helix (bHLH) transcription factor necessary for oligodendrocyte and motor neuron differentiation and development in the CNS (12,13). Olig2 has been shown to be a useful marker for primary and mature oligodendrocytes and is universally expressed in diffuse gliomas (14,15).

  1. Bradl, M. and Lassmann, H. (2010) Acta Neuropathol 119, 37-53.
  2. Harauz, G. and Boggs, J.M. (2013) J Neurochem 125, 334-61.
  3. Kozlov, G. et al. (2003) J Biol Chem 278, 46021-8.
  4. Lee, J. et al. (2005) J Cell Biol 170, 661-73.
  5. Thomson, C.E. et al. Dev Neurosci 27, 27-36.
  6. Quarles, R.H. (2007) J Neurochem 100, 1431-48.
  7. Nguyen, T. et al. (2009) J Neurosci 29, 630-7.
  8. Karram, K. et al. (2005) J Anat 207, 735-44.
  9. Küspert, M. et al. (2011) Nucleic Acids Res 39, 1280-93.
  10. Stolt, C.C. et al. (2002) Genes Dev 16, 165-70.
  11. Johns, T.G. and Bernard, C.C. (1999) J Neurochem 72, 1-9.
  12. Zhou, Q. et al. (2000) Neuron 25, 331-43.
  13. Takebayashi, H. et al. (2002) Curr Biol 12, 1157-63.
  14. Ligon, K.L. et al. (2004) J Neuropathol Exp Neurol 63, 499-509.
  15. Yokoo, H. et al. (2004) Am J Pathol 164, 1717-25.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    XP is a registered trademark of Cell Signaling Technology, Inc.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

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    Revision 1
    #38289

    Oligodendrocyte Marker Antibody Sampler Kit

    Oligodendrocyte Marker Antibody Sampler Kit: Image 1 Expand Image
    Confocal immunofluorescent analysis of fixed frozen mouse subiculum from wild-type (left) or an amyloid mouse model of Alzheimer's disease (right) using PLP1 (E9V1N) Rabbit mAb (green). After blocking free secondary antibody binding sites with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, the tissue was then labeled using β-Amyloid (D54D2) XP® Rabbit mAb (Alexa Fluor® 594 Conjugate) #35363 (red) and ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
    Oligodendrocyte Marker Antibody Sampler Kit: Image 2 Expand Image
    Confocal immunofluorescent analysis of fixed frozen mouse hippocampus from wild-type (left) or an amyloid mouse model of Alzheimer's disease (right) using PLP1 (E9V1N) Rabbit mAb (green). After blocking free secondary antibody binding sites with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, the tissue was then labeled using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb (Alexa Fluor® 555 Conjugate) (red) and ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
    Oligodendrocyte Marker Antibody Sampler Kit: Image 3 Expand Image
    Western blot analysis of extracts from various tissues using PLP1 (E9V1N) Rabbit mAb (upper) and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower). The upper band corresponds to the full-length PLP1 while the lower band is an alternative splice from PLP1 known as DM20.
    Oligodendrocyte Marker Antibody Sampler Kit: Image 4 Expand Image
    Western blot analysis of extracts from various cell lines using NG2/CSPG4 (E3B3G) XP® Rabbit mAb (upper) and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower). As expected, NG2/CSPG4 protein is not expressed in MCF7 cells.
    Oligodendrocyte Marker Antibody Sampler Kit: Image 5 Expand Image
    Western blot analysis of extracts from various brain tissues using CNPase (D83E10) XP® Rabbit mAb.
    Oligodendrocyte Marker Antibody Sampler Kit: Image 6 Expand Image
    Immunohistochemical analysis of paraffin-embedded human oligodendroglioma using Olig2 (E6G6Q) XP® Rabbit mAb.
    Oligodendrocyte Marker Antibody Sampler Kit: Image 7 Expand Image
    Immunoprecipitation of Olig2 protein from mouse brain tissue extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Olig2 (E6G6Q) XP® Rabbit mAb. Western blot analysis was performed using Olig2 (E6G6Q) XP® Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
    Oligodendrocyte Marker Antibody Sampler Kit: Image 8 Expand Image
    Western blot analysis of extracts from mouse, rat, and human brain tissue using Olig2 (E6G6Q) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
    Oligodendrocyte Marker Antibody Sampler Kit: Image 9 Expand Image
    Confocal immunofluorescent analysis of fixed frozen postnatal day five (P5) mouse hippocampus, labeled with Olig2 (E6G6Q) XP® Rabbit mAb (left, red) and co-labeled with GFAP (GA5) Mouse mAb (Alexa Fluor® 488 Conjugate) #3655 (right, green) and DAPI #4083 (right, blue).
    Oligodendrocyte Marker Antibody Sampler Kit: Image 10 Expand Image
    Confocal immunofluorescent analysis of fixed frozen mouse cerebellum, labeled with Olig2 (E6G6Q) XP® Rabbit mAb (left, red) and co-labeled with GFAP (GA5) Mouse mAb (Alexa Fluor® 488 Conjugate) #3655 (right, green) and DAPI #4083 (right, blue).
    Oligodendrocyte Marker Antibody Sampler Kit: Image 11 Expand Image
    Western blot analysis of extracts from SK-MEL-28 and HeLa cells using Sox10 (E6B6I) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). As expected, HeLa is low or negative for Sox10 expression.
    Oligodendrocyte Marker Antibody Sampler Kit: Image 12 Expand Image
    After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
    Oligodendrocyte Marker Antibody Sampler Kit: Image 13 Expand Image
    Western blot analysis of extracts from mouse brain and C2C12 cells using Myelin Basic Protein (D8X4Q) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
    Oligodendrocyte Marker Antibody Sampler Kit: Image 14 Expand Image
    Western blot analysis of extracts from mouse brain, rat brain, and human cerebellum using MAG (D4G3) XP® Rabbit mAb.
    Oligodendrocyte Marker Antibody Sampler Kit: Image 15 Expand Image
    Western blot analysis of extracts from various tissues using MOG (E5K6T) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
    Oligodendrocyte Marker Antibody Sampler Kit: Image 16 Expand Image
    Confocal immunofluorescent analysis of mouse cerebellum (left) and striatum (right) using PLP1 (E9V1N) Rabbit mAb (green) and GFAP (GA5) Mouse mAb #3670 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
    Oligodendrocyte Marker Antibody Sampler Kit: Image 17 Expand Image
    Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human NG2/CSPG4 protein (hNG2/CSPG4-Myc/DDK; +), using NG2/CSPG4 (E3B3G) XP® Rabbit mAb (upper), DYKDDDDK Tag Antibody #2368 (middle), and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower).
    Oligodendrocyte Marker Antibody Sampler Kit: Image 18 Expand Image
    Confocal immunofluorescent analysis of rat cerebellum using CNPase (D83E10) XP® Rabbit mAb (green) and α/β-Synuclein (Syn205) Mouse mAb #2644 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Oligodendrocyte Marker Antibody Sampler Kit: Image 19 Expand Image
    Immunohistochemical analysis of paraffin-embedded normal human brain using Olig2 (E6G6Q) XP® Rabbit mAb.
    Oligodendrocyte Marker Antibody Sampler Kit: Image 20 Expand Image
    Immunoprecipitation of Sox10 from SK-MEL-5 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Sox10 (E6B6I) XP® Rabbit mAb. Western blot analysis was performed usingSox10 (E6B6I) XP® Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as the secondary antibody.
    Oligodendrocyte Marker Antibody Sampler Kit: Image 21 Expand Image
    Immunohistochemical analysis of a paraffin-embedded human Alzheimer's brain using Myelin Basic Protein (D8X4Q) XP® Rabbit mAb.
    Oligodendrocyte Marker Antibody Sampler Kit: Image 22 Expand Image
    Confocal immunofluorescent analysis of rat cerebellum using MAG (D4G3) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Oligodendrocyte Marker Antibody Sampler Kit: Image 23 Expand Image
    Immunohistochemical analysis of paraffin-embedded human cerebellum using MOG (E5K6T) XP® Rabbit mAb.
    Oligodendrocyte Marker Antibody Sampler Kit: Image 24 Expand Image
    Western blot analysis of extracts from HeLa cells, either wild-type (+/+) or CSPG4 knockout (-/-), using NG2/CSPG4 (E3B3G) XP® Rabbit mAb (upper) and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower).
    Oligodendrocyte Marker Antibody Sampler Kit: Image 25 Expand Image
    Immunohistochemical analysis of paraffin-embedded human appendix using Olig2 (E6G6Q) XP® Rabbit mAb.
    Oligodendrocyte Marker Antibody Sampler Kit: Image 26 Expand Image
    Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using Sox10 (E6B6I) XP® Rabbit mAb performed on the Leica® BOND Rx.
    Oligodendrocyte Marker Antibody Sampler Kit: Image 27 Expand Image
    Immunohistochemical analysis of a paraffin-embedded mouse brain using Myelin Basic Protein (D8X4Q) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
    Oligodendrocyte Marker Antibody Sampler Kit: Image 28 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse brain using MOG (E5K6T) XP® Rabbit mAb.
    Oligodendrocyte Marker Antibody Sampler Kit: Image 29 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon carcinoma using NG2/CSPG4 (E3B3G) XP® Rabbit mAb.
    Oligodendrocyte Marker Antibody Sampler Kit: Image 30 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Olig2 (E6G6Q) XP® Rabbit mAb.
    Oligodendrocyte Marker Antibody Sampler Kit: Image 31 Expand Image
    Immunohistochemical analysis of paraffin-embedded human ductal breast carcinoma using Sox10 (E6B6I) XP® Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
    Oligodendrocyte Marker Antibody Sampler Kit: Image 32 Expand Image
    Confocal immunofluorescent analysis of rat brain using Myelin Basic Protein (D8X4Q) XP® Rabbit mAb (green) and Synaptophysin (7H12) Mouse mAb (IF Formulated) #9020 (red). Blue pseudocolor = DRAQ5®#4084 (fluorescent DNA dye).
    Oligodendrocyte Marker Antibody Sampler Kit: Image 33 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse cerebellum (left, positive) or colon (right, negative) using MOG (E5K6T) XP® Rabbit mAb. Note the lack of staining of the myenteric plexus ganglion cells in the mouse colon.
    Oligodendrocyte Marker Antibody Sampler Kit: Image 34 Expand Image
    Immunohistochemical analysis of paraffin-embedded human oral squamous cell carcinoma using NG2/CSPG4 (E3B3G) XP® Rabbit mAb.
    Oligodendrocyte Marker Antibody Sampler Kit: Image 35 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse brain, hippocampus (left), cerebellum (middle), and a nerve fiber tract in the striatum (right) using Olig2 (E6G6Q) XP® Rabbit mAb.
    Oligodendrocyte Marker Antibody Sampler Kit: Image 36 Expand Image
    Immunohistochemical analysis of paraffin-embedded human squamous carcinoma of the tongue using Sox10 (E6B6I) XP® Rabbit mAb.
    Oligodendrocyte Marker Antibody Sampler Kit: Image 37 Expand Image
    Confocal immunofluorescent analysis of mouse primary neurons and neuroglia using Myelin Basic Protein (D8X4Q) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Oligodendrocyte Marker Antibody Sampler Kit: Image 38 Expand Image
    Immunohistochemical analysis of paraffin-embedded rat brain using MOG (E5K6T) XP® Rabbit mAb.
    Oligodendrocyte Marker Antibody Sampler Kit: Image 39 Expand Image
    Immunohistochemical analysis of paraffin-embedded human renal cell carcinoma using NG2/CSPG4 (E3B3G) XP® Rabbit mAb.
    Oligodendrocyte Marker Antibody Sampler Kit: Image 40 Expand Image
    Immunohistochemical analysis of paraffin-embedded GL261 syngeneic tumor (left, positive) and Renca syngeneic tumor (right, negative) using Olig2 (E6G6Q) XP® Rabbit mAb.
    Oligodendrocyte Marker Antibody Sampler Kit: Image 41 Expand Image
    Immunohistochemical analysis of paraffin-embedded human melanoma using Sox10 (E6B6I) XP® Rabbit mAb.
    Oligodendrocyte Marker Antibody Sampler Kit: Image 42 Expand Image
    Immunohistochemical analysis of paraffin-embedded human gastrointestinal stromal tumor (left), normal liver (middle), or normal lung (right), using NG2/CSPG4 (E3B3G) XP® Rabbit mAb (top) or NG2/CSPG4 Rabbit mAb (bottom). These two antibodies detect independent, unique epitopes on human NG2/CSPG4. The similar staining patterns obtained with both antibodies help to confirm the specificity of the staining.
    Oligodendrocyte Marker Antibody Sampler Kit: Image 43 Expand Image
    Immunohistochemical analysis of paraffin-embedded GL261 syngeneic tumor using Olig2 (E6G6Q) XP® Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
    Oligodendrocyte Marker Antibody Sampler Kit: Image 44 Expand Image
    Immunohistochemical analysis of paraffin-embedded SK-MEL-5 cell pellet (left, positive) or HeLa cell pellet (right, negative) using Sox10 (E6B6I) XP® Rabbit mAb.
    Oligodendrocyte Marker Antibody Sampler Kit: Image 45 Expand Image
    Immunohistochemical analysis of paraffin-embedded normal human kidney using NG2/CSPG4 (E3B3G) XP® Rabbit mAb.
    Oligodendrocyte Marker Antibody Sampler Kit: Image 46 Expand Image
    Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using Sox10 (E6B6I) XP® Rabbit mAb.
    Oligodendrocyte Marker Antibody Sampler Kit: Image 47 Expand Image
    Immunohistochemical analysis of paraffin-embedded human B-cell non-Hodgkin's lymphoma using NG2/CSPG4 (E3B3G) XP® Rabbit mAb.
    Oligodendrocyte Marker Antibody Sampler Kit: Image 48 Expand Image
    Immunohistochemical analysis of paraffin-embedded A-375 cell pellet (left, positive) or MCF7 cell pellet (right, negative) using NG2/CSPG4 (E3B3G) XP® Rabbit mAb.
    Oligodendrocyte Marker Antibody Sampler Kit: Image 49 Expand Image
    Confocal immunofluorescent analysis of A-375 cells (left, positive) or MCF7 cells (right, negative) using NG2/CSPG4 (E3B3G) XP® Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red). Sections were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).