Revision 1

#42135Store at -20C

1 Kit

(9 x 20 microliters)

Cell Signaling Technology

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

Support: 877-678-TECH (8324)

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For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
Myelin Basic Protein (E9P7U) Mouse mAb 83683 20 µl 12-18 kDa Mouse IgG2b kappa
PLP1 (E9V1N) Rabbit mAb 28702 20 µl 20-30 kDa Rabbit IgG
MOG (E5K6T) XP® Rabbit mAb 96457 20 µl 46, 35, 28, 23 kDa Rabbit IgG
MAG (D4G3) XP® Rabbit mAb 9043 20 µl 100 kDa Rabbit IgG
Caspr (D8I3V) Rabbit mAb 97736 20 µl 190 kDa Rabbit IgG
Caspr2 (D6S1O) Rabbit mAb 61962 20 µl 150 kDa Rabbit IgG
Neurofascin 155 (D7B6O) Rabbit mAb 15035 20 µl 140-155 kDa Rabbit IgG
Neurofascin 186 (D6G6O) Rabbit mAb 15034 20 µl 200 kDa Rabbit IgG
β3-Tubulin (D71G9) XP® Rabbit mAb 5568 20 µl 55 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 Demyelinating Disease Targets Antibody Sampler Kit provides an economical means of detecting the protein components of myelin sheath. The kit includes enough antibodies to perform two western blot experiments with each 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

Myelin is the insulating material surrounding neuronal axons. The function of myelin is to promote action potential propagation down the axon to the axon terminal. Myelin is formed in the central nervous system (CNS) by specialized glial cells called oligodendrocytes and by Schwann cells in the peripheral nervous system (PNS). Oligodendrocytes and Schwann cells make concentric rings, called the myelin sheath, around the axon at regular intervals. These intervals, called nodes of ranvier, are enriched in structural proteins and ion channels, which help promote action potential propagation. Several proteins are enriched in the myelin sheath and likely help mediate the tight multi-layered membranes that make up the sheath. These proteins include myelin basic protein (MBP, [1]), myelin-associated glycoprotein (MAG, [2]), myelin proteolipid protein (PLP1, [3]) and  myelin-oligodendrocyte glycoprotein (MOG, [4]). Contactin-associated protein (Caspr) 1 & 2 (5,6) and neurofascin 155 & 186 (7,8) are nodes of ranvier-associated proteins that may play roles in generating the regular intervals of myelin along the axon. Expression of several of these proteins are altered in demyelinating diseases such as multiple sclerosis (MS). Additionally, mislocalization and/or altered expression of these proteins, compared to the axonal protein β3-tubulin, may represent altered myelin function. 

  1. Harauz, G. and Boggs, J.M. (2013) J Neurochem 125, 334-61.
  2. Li, M. et al. (1996) J Neurosci Res 46, 404-14.
  3. Thomson, C.E. et al. Dev Neurosci 27, 27-36.
  4. Johns, T.G. and Bernard, C.C. (1999) J Neurochem 72, 1-9.
  5. Rios, J.C. et al. (2000) J Neurosci 20, 8354-64.
  6. Einheber, S. et al. (1997) J Cell Biol 139, 1495-506.
  7. Charles, P. et al. (2002) Curr Biol 12, 217-20.
  8. Thaxton, C. et al. (2011) Neuron 69, 244-57.

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.

    Limited Uses

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    Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

    Revision 1
    #42135

    Demyelinating Disease Targets Antibody Sampler Kit

    Demyelinating Disease Targets 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).
    Demyelinating Disease Targets 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).
    Demyelinating Disease Targets Antibody Sampler Kit: Image 3 Expand Image
    Western blot analysis of extracts from mouse brain, mouse lung, and rat brain using Neurofascin 186 (D6G6O) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
    Demyelinating Disease Targets Antibody Sampler Kit: Image 4 Expand Image
    Western blot analysis of extracts from mouse brain, mouse lung, and rat brain using Neurofascin 155 (D7B6O) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
    Demyelinating Disease Targets Antibody Sampler Kit: Image 5 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.
    Demyelinating Disease Targets Antibody Sampler Kit: Image 6 Expand Image
    Western blot analysis of HeLa and human cerebellum using β3-Tubulin (D71G9) XP® Rabbit mAb.
    Demyelinating Disease Targets Antibody Sampler Kit: Image 7 Expand Image
    Western blot analysis of extracts from mouse and rat brain using Caspr2 (D6S1O) Rabbit mAb.
    Demyelinating Disease Targets Antibody Sampler Kit: Image 8 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.
    Demyelinating Disease Targets Antibody Sampler Kit: Image 9 Expand Image
    Western blot analysis of extracts from various tissues and cell lines using Myelin Basic Protein (E9P7U) Mouse mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
    Demyelinating Disease Targets Antibody Sampler Kit: Image 10 Expand Image
    Western blot analysis of extracts from mouse brain, rat brain, and human cerebellum using MAG (D4G3) XP® Rabbit mAb.
    Demyelinating Disease Targets Antibody Sampler Kit: Image 11 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).
    Demyelinating Disease Targets Antibody Sampler Kit: Image 12 Expand Image
    Western blot analysis of extracts from mouse brain, mouse pancreas, human cerebellum, and human pancreas using Caspr (D8I3V) Rabbit mAb (Upper) and α-Actinin (D6F6) XP® Rabbit mAb (HRP Conjugate) #12413 (Lower).
    Demyelinating Disease Targets Antibody Sampler Kit: Image 13 Expand Image
    Immunoprecipitation of Neurofascin 186 from mouse brain extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Neurofascin 186 (D6G6O) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Neurofascin 186 (D6G6O) Rabbit mAb.
    Demyelinating Disease Targets Antibody Sampler Kit: Image 14 Expand Image
    Confocal immunofluorescent analysis of mouse sciatic nerve using Neurofascin 186 (D6G6O) Rabbit mAb (left, green) or Neurofascin 155 (D7B6O) Rabbit mAb # 15035 (right, green). Neurofilament-L protein was labeled with Neurofilament-L (DA2) Mouse mAb #2835 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Demyelinating Disease Targets Antibody Sampler Kit: Image 15 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).
    Demyelinating Disease Targets Antibody Sampler Kit: Image 16 Expand Image
    Confocal immunofluorescent analysis of mouse cerebellum using β3-Tubulin (D71G9) XP® Rabbit mAb (green) and Tau (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Demyelinating Disease Targets Antibody Sampler Kit: Image 17 Expand Image
    Confocal immunofluorescent analysis of mouse brain using Myelin Basic Protein (E9P7U) Mouse mAb (green) and PDGF Receptor α (D1E1E) XP® Rabbit mAb #3174 (red). Samples were mounted in Prolong® Gold Antifade Reagent with DAPI #8961 (blue).
    Demyelinating Disease Targets Antibody Sampler Kit: Image 18 Expand Image
    Confocal immunofluorescent analysis of rat cerebellum using MAG (D4G3) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Demyelinating Disease Targets Antibody Sampler Kit: Image 19 Expand Image
    Immunohistochemical analysis of paraffin-embedded human cerebellum using MOG (E5K6T) XP® Rabbit mAb.
    Demyelinating Disease Targets Antibody Sampler Kit: Image 20 Expand Image
    Confocal immunofluorescent analysis of mouse sciatic nerve using Caspr (D8I3V) Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Demyelinating Disease Targets Antibody Sampler Kit: Image 21 Expand Image
    Confocal immunofluorescent analysis of mouse sciatic nerve using Neurofascin 186 (D6G6O) Rabbit mAb (left, green) or Neurofascin 155 (D7B6O) Rabbit mAb # 15035 (right, green). Neurofilament-L protein was labeled with Neurofilament-L (DA2) Mouse mAb #2835 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Demyelinating Disease Targets Antibody Sampler Kit: Image 22 Expand Image
    Confocal immunofluorescent analysis of rat spinal cord using Neurofascin 186 (D6G6O) Rabbit mAb (left, green) or Neurofascin 155 (D7B6O) Rabbit mAb # 15035 (right, green). β3-Tubulin protein was labeled with β3-Tubulin (TU-20) Mouse mAb #4466 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Demyelinating Disease Targets Antibody Sampler Kit: Image 23 Expand Image
    Confocal immunofluorescent analysis of P19 cells that were differentiated with retinoic acid, using β3-Tubulin (D71G9) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5 #4084® (fluorescent DNA dye).
    Demyelinating Disease Targets Antibody Sampler Kit: Image 24 Expand Image
    Confocal immunofluorescent analysis of mouse hippocampus (left, zoomed out) or (right, zoomed in) using Myelin Basic Protein (E9P7U) Mouse mAb (green, mouse IgG2b) and Synaptophysin (7H12) Mouse mAb (IF Formulated) #9020 (red, mouse IgG1). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
    Demyelinating Disease Targets Antibody Sampler Kit: Image 25 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse brain using MOG (E5K6T) XP® Rabbit mAb.
    Demyelinating Disease Targets Antibody Sampler Kit: Image 26 Expand Image
    Confocal immunofluorescent analysis of rat spinal cord using Neurofascin 186 (D6G6O) Rabbit mAb (left, green) or Neurofascin 155 (D7B6O) Rabbit mAb # 15035 (right, green). β3-Tubulin protein was labeled with β3-Tubulin (TU-20) Mouse mAb #4466 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Demyelinating Disease Targets Antibody Sampler Kit: Image 27 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.
    Demyelinating Disease Targets Antibody Sampler Kit: Image 28 Expand Image
    Immunoprecipitation of β3-Tubulin 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 β3-Tubulin (D71G9) XP® Rabbit mAb. Western blot analysis was performed using β3-Tubulin (D71G9) XP® Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb #3678 was used as the secondary antibody.
    Demyelinating Disease Targets Antibody Sampler Kit: Image 29 Expand Image
    Immunohistochemical analysis of paraffin-embedded rat brain using MOG (E5K6T) XP® Rabbit mAb.