Catalogue# RPCA-NF-H: Rabbit Polyclonal Antibody to Neurofiilament NF-H
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, a-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. To raise this antibody bovine intermediate filaments were prepared from spinal cords by the method of Delacourte et al. and the cytoskeletal material was dissolved in 6M urea. Individual neurofilament subunits were purified by ion exchange chromatography on DEAE cellulose followed by preparative gel electrophoresis. The form of NF-H isolated is the heavily phosphorylated axonal variant, and the antibody generated recognizes predominantly axonal neurofilaments. The HGNC name for this protein is NEFH.
Left: Lane labelled CBB shows Coomassie Brilliant Blue stained crude extract of rat spinal cord, with the prominent major neurofilament subunit indicated; In rodents, NF-H runs at 200kDa, NF-M at 145kDa and NF-L at 68kDa. The Rab lane shows a blot probed with rabbit anti NF-H antibody RPCA-NF-H. A prominent band at an apparent SDS-PAGE molecular weight of 200kDa corresponds to phosphorylated form of NF-H. The lane marked Rab shows a similar blot probed with our rabbit antibody to NF-H, CPCA-NF-H. Right: shows a section of rat spinal cord stained with RPCA-NF-H (green) and EnCor's new monoclonal antibody to ubiquitin C-terminal hydrolase 1 (UCHL1) clone MCA-BH7 (red). The neurofilament NF-H antibody binds primarily to phosphorylated axonal forms of NF-H, and so stains axons coursing between the large UCHL1 positive neurons. These large cells are a-motorneurons and UCHL1 protein is a major component of the perikarya and dendrites of these cells.
Antibody Characteristics: Antibody was raised in rabbit against purified bovine NF-H. Store at 4°C or -20°C. Avoid repeat freezing and thawing.
Suggestions for use: We suggest a dilution of 1:1,000 for immunofluorescence microscopy and 1:10,000 for western blots. The antibody has also been used successfully in the detection role in ELISA (1).
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References:
1. Shaw G, Yang C, Ellis R, Anderson K, Parker Mickle J, Scheff S, Pike B, Anderson DK and Howland DR. Hyperphosphorylated neurofilament NF-H is a serum biomarker of axonal injury. Biochem Biophys Res Commun. 336:1268-1277 (2005).
Limitations: This product is for research use only and is not approved for use in humans or in clinical diagnosis.
Availability and Price: Available for shipping now, $200 US per aliquot of 100ml of crude serum, enough for hundreds of experiments. For order form press here
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