Anti-Neurofilament M/NEFM Antibody, clone 2H3

Neurofilament medium polypeptide (UniProt: P12839; also known as NF-M, 160 kDa neurofilament protein, Neurofilament 3, Neurofilament triplet M protein) is encoded by the Nefm (also known as Nef3, Nfm) gene (Gene ID: 24588) in rat. Neurofilaments are intermediate filaments 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. They are built from three intertwined protofibrils, which themselves are composed of two tetrameric protofilament complexes 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. Neurofilaments contain a number of repeats of the tripeptide KSP (Lys-Ser-Pro) and NF-M is phosphorylated on a number of the serine residues in this motif by protein kinase A and C. Phosphorylation of NF-M results in the formation of interfilament cross bridges that are important in the maintenance of axonal caliber. Deletion of the phosphorylated tail domain of NF-M is reported to inhibit radial growth of axons and reduce their conduction velocities. However, mice expressing NF-M subunits that lack phosphorylation at KSP sites along the tail domain have normal axon calibers and conduction velocities, indicating that the NF-M tail domain, but not tail phosphorylation is crucial for axon radial growth. The level of phosphorylation is shown to be altered developmentally and coincides with a change in the neurofilament function. Mutations in the rod domain region of NF-M have been identified in early onset Parkinson s disease. Also, in the brains of Alzheimer s disease patients, a reciprocal relationship between O-GlcNAcylation and phosphorylation of NF-M has been reported where reduced O-GlcNAcylation is accompanied with increased KSP phosphorylation in NF-M. (Ref.: Yuan, A., et al. (2017). Cold Spring Harb. Perspect. Biol. 9(4); a018309; Yuan, A., et al. (2012). J. Cell Sci. 125(14); 3257-3263).

Membrane preparations from E14-E15 rat brain tissue.

Quality Control Testing

Evaluated by Western Blotting in Rat brain tissue extracts.

Western Blotting Analysis: A 1:500 dilution of this antibody detected Neurofilament medium polypeptide (NF-M) in Rat brain tissue extract.

Tested Applications

Western Blotting Analysis: A 1:500 dilution from a representative lot detected Neurofilament medium polypeptide (NF-M) in mouse brain tissue extract.

Immunohistochemistry Applications: A representative lot detected Neurofilament medium polypeptide (NF-M) in Immunohistochemistry applications (Clugston, R.D., et al. (2010). Am J Respir Cell Mol Biol. 42(3):276-85; Lysakowski, A., et al. (2011). J Neurosci. 31(27):10101-14; Kridsada, K., et al. (2018). Cell Rep. 23(10):2928-2941; Latremoliere, A., et al. (2018). Cell Rep. 24(7):1865-1879.e.9).

Immunocytochemistry Analysis: A representative lot detected Neurofilament medium polypeptide (NF-M) in Immunocytochemistry applications (Latremoliere, A., et al. (2018). Cell Rep. 24(7):1865-1879.e.9).

Immunofluorescence Analysis: A representative lot detected Neurofilament medium polypeptide (NF-M) in Immunofluorescence applications (Latremoliere, A., et al. (2018). Cell Rep. 24(7):1865-1879.e.9).

Western Blotting Analysis: A representative lot detected Neurofilament medium polypeptide (NF-M) in Western Blotting applications (Fernandez-Cerado, C., et al. (2021). J Neural Transm (Vienna). 128(4):575-587).

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.

Clone 2H3 is a mouse monoclonal antibody that detects Neurofilament-M (NEFM)

Purified mouse monoclonal antibody IgG1 in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

0.5 mg/mL. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Recommended storage: +2°C to +8°C.

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

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