Revision 1

#9781Store at -20C

1 Kit

(4 x 20 microliters)

Cell Signaling Technology

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

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
Neurofilament-M (RMO 14.9) Mouse mAb 2838 20 µl 160 kDa Mouse IgG1
Neurofilament-L (DA2) Mouse mAb 2835 20 µl 70 kDa Mouse IgG1
Neurofilament-L (C28E10) Rabbit mAb 2837 20 µl 70 kDa Rabbit IgG
Neurofilament-H (RMdO 20) Mouse mAb 2836 20 µl 180-220 kDa Mouse IgG1
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 
Anti-mouse IgG, HRP-linked Antibody 7076 100 µl Horse 

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

Description

The Neurofilament Antibody Sampler Kit provides an economical means of evaluating neurofilaments. The kit contains enough primary and secondary antibodies to perform 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 cytoskeleton consists of three types of cytosolic fibers: actin microfilaments, 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). Similar in structure to other intermediate filament proteins, neurofilaments have a globular amino-terminal head, a central α-helical rod domain, and a carboxy-terminal tail. A heterotetrameric unit (NFL-NFM and NFL-NFH) forms a protofilament, with eight protofilaments comprising the typical 10 nm intermediate filament (2). While neurofilaments are critical for radial axon growth and determine axon caliber, microtubules are involved in axon elongation. PKA phosphorylates the head domain of NFL and NFM to inhibit neurofilament assembly (3,4). Research studies have shown neurofilament accumulations in many human neurological disorders, including Parkinson's disease (in Lewy bodies along with α-synuclein), Alzheimer's disease, Charcot-Marie-Tooth disease, and Amyotrophic Lateral Sclerosis (ALS) (1).

  1. Al-Chalabi, A. and Miller, C.C. (2003) Bioessays 25, 346-55.
  2. Cohlberg, J.A. et al. (1995) J Biol Chem 270, 9334-9.
  3. Hisanaga, S. et al. (1994) Mol Biol Cell 5, 161-72.
  4. Sihag, R.K. et al. (1999) J Neurochem 72, 491-9.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a 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
    #9781

    Neurofilament Antibody Sampler Kit

    Neurofilament Antibody Sampler Kit: Image 1 Expand Image
    Simple WesternTM analysis of lysates (0.1 mg/mL) from Mouse Brain cells using Neurofilament-H (RMdO20) Mouse mAb #2836. 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 JessTM Simple Western instrument from ProteinSimple, a BioTechne brand, using the 66-440 kDa separation module.
    Neurofilament Antibody Sampler Kit: Image 2 Expand Image
    Western blot analysis of extracts from rat brain, mouse brain, U87-MG cells, C6 cells and human cerebellum, using Neurofilament-L (DA2) Mouse mAb.
    Neurofilament Antibody Sampler Kit: Image 3 Expand Image
    Western blot analysis of extracts from mouse brain, rat brain or CAD cells, using Neurofilament-H (RMdO 20) Mouse mAb.
    Neurofilament Antibody Sampler Kit: Image 4 Expand Image
    Western blot analysis of extracts from mouse brain, HeLa cells and rat brain, using Neurofilament-L (C28E10) Rabbit mAb.
    Neurofilament Antibody Sampler Kit: Image 5 Expand Image
    Western blot analysis of extracts from rat brain, untreated or treated with lambda phosphatase, using Neurofilament-M (RMO 14.9) Mouse mAb.
    Neurofilament Antibody Sampler Kit: Image 6 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.
    Neurofilament Antibody Sampler Kit: Image 7 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.
    Neurofilament Antibody Sampler Kit: Image 8 Expand Image
    Immunohistochemical analysis of paraffin-embedded human brain using Neurofilament-L (DA2) Mouse mAb.
    Neurofilament Antibody Sampler Kit: Image 9 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse brain using Neurofilament-L (C28E10) Rabbit mAb.
    Neurofilament Antibody Sampler Kit: Image 10 Expand Image
    Immunohistochemical analysis of paraffin-embedded human medulloblastoma, using Neurofilament-L (DA2) Mouse mAb.
    Neurofilament Antibody Sampler Kit: Image 11 Expand Image
    Confocal immunofluorescent image of mouse cerebellum labeled with Neurofilament-H (RMdO 20) Mouse mAb (green) and Calbindin Antibody #2136 (red). Blue pseudocolor = DRAQ5 (fluorescent DNA dye).
    Neurofilament 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).
    Neurofilament Antibody Sampler Kit: Image 13 Expand Image
    Confocal immunofluorescent image of mouse cerebellum labeled with Neurofilament-L (DA2) Mouse mAb (green) and CREB (48H2) Rabbit mAb #9197 (red). Blue pseudocolor = DRAQ5® (fluorescent DNA dye).
    Neurofilament 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).