Revision 1

#8572Store at -20C

1 Kit

(5 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
GFAP (D1F4Q) XP® Rabbit mAb 12389 20 µl 50 kDa Rabbit IgG
CNPase (D83E10) XP® Rabbit mAb 5664 20 µl 47 kDa Rabbit IgG
β3-Tubulin (D71G9) XP® Rabbit mAb 5568 20 µl 55 kDa Rabbit IgG
Nestin (Rat-401) Mouse mAb 4760 20 µl Mouse IgG1
Neurofilament-L (C28E10) Rabbit mAb 2837 20 µl 70 kDa Rabbit IgG

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

Description

The Neuronal Marker IF Antibody Sampler Kit provides an economical means for labeling neuronal structures by immunofluorescence (IF-F). This kit includes enough primary antibody to perform at least forty IF-F 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 antibody.

Background

The antibodies in this kit serve as neuronal markers to determine protein localization in neurons. The cytoskeleton consists of three types of cytosolic fibers: microfilaments (actin filaments), intermediate filaments, and microtubules. Neurofilaments are the major intermediate filaments found in neurons and consist of light (NFL), medium (NFM), and heavy (NFH) subunits (1). Nestin is an intermediate filament family member protein that is structurally related to the neurofilament proteins (2). Globular tubulin subunits comprise the microtubule building block, with α/β-tubulin heterodimers forming the tubulin subunit common to all eukaryotic cells (3). High CNPase expression is seen in oligodendrocytes and Schwann cells as CNPase accounts for roughly 4% of the total myelin protein in the central nervous system (4). CNPase binds to tubulin heterodimers and plays a role in tubulin polymerization and oligodendrocyte process outgrowth (5). GFAP filaments are characteristic of differentiated and mature brain astrocytes. Thus, GFAP is commonly used by investigators as a marker for intracranial and intraspinal tumors arising from astrocytes (6).

  1. Al-Chalabi, A. and Miller, C.C. (2003) Bioessays 25, 346-55.
  2. Michalczyk, K. and Ziman, M. (2005) Histol Histopathol 20, 665-71.
  3. Westermann, S. and Weber, K. (2003) Nat Rev Mol Cell Biol 4, 938-47.
  4. Kozlov, G. et al. (2003) J Biol Chem 278, 46021-8.
  5. Lee, J. et al. (2005) J Cell Biol 170, 661-73.
  6. Goebel, H.H. et al. (1987) Acta Histochem Suppl 34, 81-93.

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.
    U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    Limited Uses

    Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

    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
    #8572

    Neuronal Marker IF Antibody Sampler Kit

    Neuronal Marker IF Antibody Sampler Kit: Image 1 Expand Image
    Simple Western™ analysis of tissue extract (0.1 mg/mL) from mouse brain using GFAP (D1F4Q) XP® Rabbit mAb #12389. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
    Neuronal Marker IF Antibody Sampler Kit: Image 2 Expand Image
    Western blot analysis of extracts from mouse brain, NIH/3T3 cells, rat brain, and C6 cells using GFAP (D1F4Q) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
    Neuronal Marker IF Antibody Sampler Kit: Image 3 Expand Image
    Western blot analysis of extracts from mouse brain, HeLa cells and rat brain, using Neurofilament-L (C28E10) Rabbit mAb.
    Neuronal Marker IF Antibody Sampler Kit: Image 4 Expand Image
    Immunohistochemical analysis of paraffin-embedded rat brain, P1 (left) or P14 (right), using Nestin (Rat-401) Mouse mAb (top) or Neurofilament-L (C28E10) Rabbit mAb #2837 (lower). Note decreased Nestin staining as NFL staining increases in the developing rat brain.
    Neuronal Marker IF Antibody Sampler Kit: Image 5 Expand Image
    Western blot analysis of HeLa and human cerebellum using β3-Tubulin (D71G9) XP® Rabbit mAb.
    Neuronal Marker IF Antibody Sampler Kit: Image 6 Expand Image
    Western blot analysis of extracts from various brain tissues using CNPase (D83E10) XP® Rabbit mAb.
    Neuronal Marker IF Antibody Sampler Kit: Image 7 Expand Image
    Confocal immunofluorescent analysis of rat cerebellum using GFAP (D1F4Q) XP® Rabbit mAb (green) and Neurofilament-H (RMdO 20) Mouse mAb #2836 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Neuronal Marker IF Antibody Sampler Kit: Image 8 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse brain using Neurofilament-L (C28E10) Rabbit mAb.
    Neuronal Marker IF Antibody Sampler Kit: Image 9 Expand Image
    Confocal immunofluorescent analysis of P1 rat brain (left) and adult rat brain (right) using Nestin (Rat-401) Mouse mAb (green) and Neurofilament-L (C28E10) Rabbit mAb #2837 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Neuronal Marker IF Antibody Sampler Kit: Image 10 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).
    Neuronal Marker IF Antibody Sampler Kit: Image 11 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).
    Neuronal Marker IF Antibody Sampler Kit: Image 12 Expand Image
    Immunohistochemical analysis of paraffin-embedded human brain using Neurofilament-L (C28E10) Rabbit mAb in the presence of control peptide (left) or Neurofilament-L blocking peptide #1005 (right).
    Neuronal Marker IF Antibody Sampler Kit: Image 13 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).
    Neuronal Marker IF Antibody Sampler Kit: Image 14 Expand Image
    Confocal immunofluorescent analysis of normal rat cerebellum using Neurofilament-L (C28E10) Rabbit mAb (green) and GFAP (GA5) Mouse mAb #3670 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Neuronal Marker IF Antibody Sampler Kit: Image 15 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.