Peptide mapping by CNBr fragmentation using a sodium dodecyl sulfate-polyacrylamide minigel system.
The method of peptide mapping by sodium dodecyl sulfate-gel electrophoresis following partial protein fragmentation with cyanogen bromide was adapted for a polyacrylamide minigel system. The combined use of the discontinuous gel electrophoresis system of J. P. Doucet and J. M. Trifaró [1988) Anal. Biochem. 168, 265-271) and a vertical polyacrylamide minigel system produced the following advantages over other procedures: (a) the ability to resolve cyanogen bromide cleavage fragments over a broad molecular mass range while yielding very sharp protein staining bands; (b) well-defined peptide maps are produced with as little as 2 micrograms of protein; (c) less time is required to perform fragmentation with cyanogen bromide, to equilibrate the gel slices in sodium dodecyl sulfate buffer, as well as to perform the electrophoresis; and (d) the cyanogen bromide fragmentation patterns are highly reproducible.
Plaxton
W C
WC
Department of Biology, Queen's University, Kingston, Ontario, Canada.
Moorhead
G B
GB
D003488:Cyanogen Bromide; D004591:Electrophoresis, Polyacrylamide Gel; D007202:Indicators and Reagents; D007700:Kinetics; D010449:Peptide Mapping / Q000379:methods*; D012967:Sodium Dodecyl Sulfate
D007202:Indicators and Reagents; D012967:Sodium Dodecyl Sulfate; D003488:Cyanogen Bromide