Merck
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MAB1615

Sigma-Aldrich

Anti-Neurofilament 70 kDa Antibody, clone DA2

culture supernatant, clone DA2, Chemicon®

Synonym(s):
Anti-CMTDIG, Anti-NF-L, Anti-NF68, Anti-CMT1F, Anti-PPP1R110, Anti-NFL
eCl@ss:
32160702
NACRES:
NA.41

Quality Level

biological source

mouse

antibody form

culture supernatant

antibody product type

primary antibodies

clone

DA2, monoclonal

species reactivity

bovine, human, pig, rat, mouse

manufacturer/tradename

Chemicon®

technique(s)

immunocytochemistry: suitable
immunohistochemistry: suitable
western blot: suitable

isotype

IgG1

NCBI accession no.

UniProt accession no.

shipped in

dry ice

Gene Information

human ... NEFL(4747)

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

Linkage

Replaces: 04-1112

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.

Legal Information

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

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.

Storage Class Code

10 - Combustible liquids

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Certificate of Analysis

Enter Lot Number to search for Certificate of Analysis (COA).

Certificate of Quality

Enter Lot Number to search for Certificate of Quality (COQ).

Cytoarchitecture of the mouse neocortex revealed by the low-molecular-weight neurofilament protein subunit.
Melissa Paulussen,Sandy Jacobs,Estelle Van der Gucht,Patrick R Hof,Lutgarde Arckens
Brain Structure &Amp; Function null
Specific human astrocyte subtype revealed by affinity purified GFAP antibody; unpurified serum cross-reacts with neurofilament-L in Alzheimer.
Middeldorp, J; van den Berge, SA; Aronica, E; Speijer, D; Hol, EM
Testing null
Renata Novotny et al.
The Journal of neuroscience : the official journal of the Society for Neuroscience, 36(18), 5084-5093 (2016-05-06)
The aggregation of amyloid-β peptide (Aβ) in brain is an early event and hallmark of Alzheimer's disease (AD). We combined the advantages of in vitro and in vivo approaches to study cerebral β-amyloidosis by establishing a long-term hippocampal slice culture
Carbon filament implants promote axonal growth across the transected rat spinal cord.
Khan, T, et al.
Brain Research, 541, 139-145 (1991)
T Nishio et al.
Neuroscience, 155(1), 90-103 (2008-06-24)
Following transection of the spinal cord, severed axonal ends retract from the lesion site and attempt regeneration within 24 h of injury. Molecular mechanisms underlying such rapid axonal reactions after severance are not fully characterized so far. To better understand

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