Neurofilaments are the 10nm or intermediate filament proteins found specifically in neurons, and are composed predominantly of four major proteins called NF-L, NF-M, NF-H and alpha-internexin. NF-H is the neurofilament heavy or high molecular weight polypeptide and runs on SDS-PAGE gels at 200-220kD in the heavily phosphorylated axonal form. The molecule has an unusual and interesting region consisting of multiple Lysine-Serine-Proline peptides, about 40 of these in human. These peptide repeats are heavily phosphorylated on the Serine residues in axons. Enzymatic removal of these phosphate groups will increase the SDS-PAGE mobility to about 160kD, likely due to protein conformational changes due to the removal of charge. Even the non-phosphorylated form runs aberrantly on SDS-PAGE, as the real molecular weight of NF-H is about 110kD, with some variation in different species. This is likely due to an unusually high content of charged amino acids. Non-phosphorylated forms of NF-H are found in dendrites and perikarya and early in development, but the majority of NFH in the adult is this heavily phosphorylated axonal form. Source: Isolated from pig spinal cord using a modification of the method of Leung and Liem (ref. 3), which purifies out the heavily phosphorylated axonal form, often referred to as pNF-H. Applications: Suitable for use in Western Blot and ELISA. Other applications have not been tested. Recommended Dilutions: Optimal dilutions to be determined by the researcher. Storage and Stability: May be stored at 4C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20C. Aliquots are stable for 6 months after receipt at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
Molekulargewicht:
220
Reinheit:
Purified
Formulierung:
Supplied as a liquid in 10mM phosphate, pH 7.5, 6M urea.
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