Neurofilaments are the 10 nm or intermediate filament proteins found specifically in neurons, and are composed predominantly of four major proteins called NF-L, NF-M, NF-H and α-internexin. NF-L is the neurofilament light or low molecular weight polypeptide and runs on SDS-PAGE gels at about 68 kDa.
A cDNA encoding full length human neurofilament NF-L was inserted into an 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 is diluted to 1 mg/mL and is supplied in 6M urea.
Image: Coomassie Brilliant Blue stained SDS-PAGE gel of various recombinant proteins. His-tagged human Neurofilament NF-L, was expressed and purified from E. coli BL21 using immobilized metal affinity chromatography. 1µg of protein was run on each lane, and the lane indicated with "NF-L" contains the Neurofilament NF-L protein. The other lanes show recombinant His-tagged Peripherin, α-Internexin and Vimentin as indicated. Protein molecular weight standards are in the first lane and apparent molecular weights are as indicated.
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