Catalogue RPCA-NF-L: Polyclonal Antibody to Neurofilament Subunit NF-L
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 purified NF-L and recognizes all mammalian and avian NF-L molecules.
Left: Western blot of whole rat cerebellum homogenate stained with RPCA-NF-L, at dilution of 1:20,000. A prominent band running with an apparent SDS-PAGE molecular weight of ~68kDa corresponds to rodent NF-L. Human, cow and bovine NF-L run a little slower, at about 70kDa. Right: Mixed neuron/glia cultures from newborn rat brain stained with MCA-7C5 antibody to peripherin (green) and rabbit polyclonal antibody to NF-L RPCA-NF-L (red channel). A class of large neurons, like the one in the middle of this image, contain peripherin, while the majority of neurons and their processes contain NF-L and not peripherin. Interestingly, the periperin positive cells often contain a cytoplasmic inclusion next to the nucleus which stains for both peripherin and NF-L, and so appears golden in this kind of image. The blue channel reveals the localization of DNA
Antibody Characteristics: To raise this antibody pig intermediate filaments were prepared from spinal cords by the method of Delacourte et al. and this cytoskeletal material was dissolved in 6M urea. NF-L was purified by ion exchange chromatography, followed by preparative gel electrophoresis. 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.
Price and Availability: - We currently supply 100 microliter aliquots for $200. Material is in stock and ready for immediate shipping.
Limitations: This product is for research use only and is not approved for use in humans or in clinical diagnosis.
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