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MAB1615 Sigma-Aldrich

Anti-Neurofilament 70 kDa Antibody, clone DA2

This Anti-Neurofilament 70 kDa Antibody, clone DA2 is validated for use in WB, IC, IH for the detection of Neurofilament 70 kDa.

  •  eCl@ss 32160702

  •  NACRES NA.41

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Properties

Related Categories Alphabetical Index, Antibodies, NE-NI, Primary Antibodies
clone   DA2, monoclonal
biological source   mouse
application(s)   immunocytochemistry: suitable
  immunohistochemistry: suitable
  western blot: suitable
species reactivity   bovine, human, pig, rat, mouse
shipped in   dry ice
antibody form   culture supernatant
isotype   IgG1
Quality Level   100
antibody product type   primary antibodies
mfr. no.   Chemicon®
NCBI accession no.   NM_006158.2
UniProt accession no.   P07196
Gene Information   human ... NEFL(4747)

Description

General description

Neurofilaments are a type of intermediate filament that serve as major elements of the cytoskeleton supporting the axon cytoplasm. They are the most abundant fibrillar components of the axon, being on average 3-10 times more frequent than axonal microtubules. Neurofilaments (10nm in dia.) are built from three intertwined protofibrils which are themselves composed of two tetrameric protofilament complexs of monomeric proteins. The neurofilament triplet proteins (68/70, 160, and 200 kDa) occur in both the central and peripheral nervous system and are usually neuron specific. The 68/70 kDa NF-L protein can self-assemble into a filamentous structure, however the 160 kDa NF-M and 200 kDa NF-H proteins require the presence of the 68/70 kDa NF-L protein to co-assemble. Neuromas, ganglioneuromas, gangliogliomas, ganglioneuroblastomas and neuroblastomas stain positively for neurofilaments. Although typically restricted to neurons, neurofilaments have been detected in paragangliomas and adrenal and extra-adrenal pheochromocytomas. Carcinoids, neuroendocrine carcinomas of the skin, and oat cell carcinomas of the lung also express neurofilaments. For more neurofilament information see Nervous System Cell Type Specific Marker chart online under the CHEMICON Technical Support section.

Specificity

Recognizes the low molecular weight (68-70 kDa) subunit of the neurofilament triplet (NF-L), the epitope is phosphate independent. Reacts with human and higher vertebrates.

Immunogen

Enzymatically dephosphorylated pig neurofilaments.

Application

Immunoblot and immunohistochemistry (frozen sections, little reactivity on formalin-fixed paraffin embedded material). Optimal working dilutions must be determined by end user.

IMMUNOHISTOCHEMISTRY PROTOCOL FOR MAB1615

This antibody has been used successfully on 30 mm, free floating, 4% paraformaldehyde fixed rat brain tissue. All steps are performed under constant agitation. Suggested protocol follows.

1) 3 x 10 minute washes in TBS (with or without 0.25% Triton).

2) Incubate for 30 minutes in TBS with 3% serum (same as host from secondary antibody).

3) Incubate primary antibody diluted appropriately in TBS with 1% serum (same as host from secondary antibody) (with or without 0.25% Triton) for 2 hours at room temperature followed by 16 hours at 4°C.

4) 3 x 10 minute washes in TBS.

5) Incubate with secondary antibody diluted appropriately in TBS with 1% serum (same as host from secondary antibody).

6) 3 x 10 minute washes in TBS.

7) ABC Elite (1:200 Vector Labs) in TBS.

8) 2 x 10 minute washes in TBS.

9) 1 x 10 minute wash in phosphate buffer (no saline).

10) DAB reaction with 0.06% NiCl added for intensification.

11) 2 x 10 minute washes in PBS.

12) 1 x 10 minute wash in phosphate buffer (no saline).

Research Category
Neuroscience

Research Sub Category
Neurofilament & Neuron Metabolism

Neuronal & Glial Markers

Physical form

Liquid. Contains no preservative.

Storage and Stability

Maintain at -20°C in undiluted aliquots for up to 12 months. Avoid repeated freeze-thaw cycles.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Linkage

Replaces: 04-1112

Safety & Documentation

Safety Information

Safety Information for this product is unavailable at this time.
Protocols & Articles

Articles

Antibody Basics

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Protocols

Western Blot Protocol | Immunoblotting Protocol

Western Blotting refers to the electrophoretic transfer of proteins from sodium dodecyl sulfate polyacrylamide gels to sheets of PVDF or nitrocellullose membrane, followed by immunodetection of prote...
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Peer-Reviewed Papers
15

References

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