Internal Standards for Molecular Weight Determinations of Proteins by Polyacrylamide Gel Electrophoresis

摘要: Abstract A new technique was devised to be used with molecular weight determinations of proteins on polyacrylamide gels. Internal standards consisting covalently linked 1-dimethylaminonaphthalene-5-sulfonyl- (DANS-) groups were prepared. DANS- derivatives insulin and monomer, dimer, trimer, tetramer, pentamer, hexamer egg white lysozyme serum albumin covered weights from 6,600 360,000. They concurrently applied in small amounts a single gel together the unknown sample. Proteins separated by electrophoresis 0.5% sodium dodecyl sulfate. Positions detected under an ultraviolet lamp as yellowish fluorescent bands. These bands are reproducible related positions obtained standard staining techniques. Even 0.2 µg DANS-protein detectable. Molecular unknowns determined interpolation their positions. This method envelope Escherichia coli. The major determined. X Y, which have been shown previous paper cell division DNA synthesis, respectively, reexamined found 50,000 44,000, respectively. One at 7500. protein showed very high arginine histidine ratio (about 11 times others) distinctly different all other higher weights. Thus, this is not subunit