Vimentin is a member of the intermediate or 10nm family of proteins
, which are abundant structural components of most eukaryotic cells. Intermediate filament proteins are divided into 5 classes, with class I and II including the epithelial keratins, class IV the neurofilaments and class V the nuclear lamins. Class III includes vimentin along with desmin, peripherin and glial fibrillary acidic protein (GFAP). Vimentin is found in most cell lines grown in tissue culture and in many cell types early in development. In adult animals vimentin is found in mesenchymal cells, lymphocytes, many kinds of glia, endothelial cells and many others.
Coomassie Brilliant Blue stained SDS-PAGE gel of various recombinant human proteins. His-tagged human vimentin, was expressed and purified from E. coli
using immobilized metal affinity chromatography. 1µg of protein was run on each lane, and the lane indicated with “Vim” contains the vimentin protein. The other lanes show recombinant His-tagged peripherin (Peri), α-internexin (α-int) and neurofilament NF-L (NF-L) as indicated. Protein molecular weight standards are in the first lane indicated with “St” and apparent molecular weights are as indicated.
A cDNA encoding full length human vimentin was inserted into a eukaryotic expression vector which adds an N-terminal in frame His-tag. This was transformed into E. coli and recombinant protein was purified in 6M urea using immobilized metal affinity chromatography. Purified protein was diluted to 1 mg/mL and is supplied in 6M urea.
1. Franke, W. W., Schmid, E., Osborn, M. and Weber, K. Different intermediate-sized filaments distinguished by immunofluorescence microscopy. Proc. Natl. Acad. Sci. USA 75:5034–5038 (1978).
2. Muller, M., Bhattacharya, S. S., Moore, T., Prescott, Q., Wedig, T., Herrmann, H., Magin, T. M. Dominant cataract formation in association with a vimentin assembly disrupting mutation. Hum. Molec. Genet. 18:1052-1057 (2009).
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