The Immunogen: Neurofilaments can be defined as the intermediate or 10nm diameter filaments found in neuronal cells. They are composed a mixture of subunits which often includes the neurofilament triplet proteins, NF-L, NF-M and NF-H. Neurofilaments may also include peripherin, alpha-internexin, nestin and in some cases vimentin. Antibodies to the various neurofilament subunits are very useful cell type markers since the proteins are quite abundant, biochemically stable. This antibody was raised against the C-terminal region of NF-M, a region highly conserved between NF-M molecules in different species. Accordingly the antibody recognizes all mammalian and avian NF-M molecules. The name NF-M refers to the fact that this is the "middle" or "medium" protein of neurofilament triplet, the other two proteins being "low" or "light" (NF-L) and "high" or "heavy" (NF-H). NF-M is sometimes also referred to as NEF3 or Neurofilament 3. The HGNC name for this protein is NEFM.

Left: Western blot of whole rat cerebellum homogenate stained with RPCA-NF-M, at dilution of 1:20,000. A prominent band running with an apparent SDS-PAGE molecular weight of ~145kDa corresponds to rodent NF-M. Human, cow and bovine NF-M run a little slower, at about 160kDa. Right: Section of rat cerebral cortex stained with RPCA-NF-M (red), which reveals the perikarya of pyramidal neurons and dendrites and axons surrounding them. The green channel shows staining with a monoclonal antibody to the beta-adrendergic receptor kinase 1.

Antibody Characteristics: Antibody was raised in rabbit which was injected with a recombinant fusion protein containind the extreme C-terminal segment of rat NF-M. The antibody is similar to that described in reference 1 below, and has been marketed by many companies for many years. Store at 4°C or -20°C. Avoid repeat freezing and thawing.

Suggestions for use: Try at dilutions of 1:500 to 1:1,000 for immunofluorescence, and 1:5,000 for ABC or other enzyme linked immunocytochemical procedures. For western blots try at 1:20,000.

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References:

1.  Harris, J., Ayyub, C. and Shaw G. A molecular dissection of the carboxyterminal tails of the major neurofilament subunits NF-M and NF-H. J Neurosci Res 30:47-62 1991.

2. Wang, Y., Zhang, J., Mori, S. and Nathans. J. Axonal Growth and Guidance Defects in Frizzled3 Knock-Out Mice: A Comparison of Diffusion Tensor Magnetic Resonance Imaging, Neurofilament Staining, and Genetically Directed Cell Labeling. J. Neurosci. 26:355-364 2006.

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