The Immunogen: Neurofilaments are the 10nm or intermediate filament proteins found specifically in neurons, and are composed predominantly of three major proteins called NF-L, NF-M and NF-H (HUGO names are NEFL, NEFM and NEFH respectively). NF-H is the neurofilament high or heavy molecular weight polypeptide and runs on SDS-PAGE gels at 200-220 kDa, with some variability across species boundaries. Antibodies to NF-H are useful for identifying neuronal cells and their processes in tissue sections and in tissue culture. NF-H antibodies can also be useful in the diagnostics of neurofilament accumulations seen in many neurological diseases, such as Amyotrophic Lateral Sclerosis (also known as Lou Gehrig's disease) and Alzheimer's disease. MCA-NAP4 is one of numerous antibodies which reacts preferentially with the axonal phosphorylated forms of NF-H. Interestingly these phosphorylated forms of NF-H are normally restricted to axons, while less phosphorylated forms are found in dendrites. However in numerous damage and disease states, phosphorylated NF-H can be detected with MCA-NAP4 in dendritic and perikaryal neurofilaments. The HGNC name for this protein is SOCS7.

Left: Rat spinal cord homogenate showing the major intermediate filament proteins of the nervous system (lane 1). The remaining lanes show blots of this material stainted with various antibodies including MCA-NAP4 (lane 2). Right: Human cerebellar cortex fixed in formalin, embedded in paraffin and stained with MCA-NAP4 using the ABC (avidin biotin conjugate) method. The section was counterstained with heamatoxylin-eosin (blue). MCA-NAP4 stains prominent basket cell axons surrounding the large Purkinje neurons. Granule cell layer at bottom of image, molecular layer at top.

Antibody characteristics: MCA-NAP4 is a mouse monoclonal antibody raised against a preparation of native NF-H purified from pig spinal cord. MCA-NAP4 is an IgG1 class antibody with a k light chain. It recognizes phosphorylated NF-H KSP (lysine-serine-proline) type sequences. In some species there is some cross-reactivity with the related phosphorylated KSP sequences found in the related neurofilament subunit NF-M. The antibody recognizes NF-H strongly in all mammals tested to date and also in chicken. It recognizes neurofilaments in frozen sections in tissue culture and in formalin fixed sections.

Suggestions for use: The ascites solution has a high titre and can be used at dilutions of at least 1:1,000 in immunofluorescence experiments. In western blotting using chemiluminescence it can be used at dilutions of 1:10,000 or lower.

Limitations: This product is for research use only and is not approved for use in humans or in clinical diagnosis.

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.

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