Enhanced removal of detergent and recovery of enzymatic activity following sodium dodecyl sulfate-polyacrylamide gel electrophoresis: use of casein in gel wash buffer.
The inclusion of 1% casein or bovine serum albumin in buffer used to reactivate enzymes subjected to sodium dodecyl sulfate (SDS)-polyacrylamide electrophoresis resulted in accelerated removal of SDS and restoration of nuclease and beta-galactosidase enzyme activities. Nuclease and beta-galactosidase activities which are absent from gels after longer wash procedures are detectable with this technique. Enzyme activity in gels prepared with SDS which contained inhibitory contaminants was partially restored by the casein wash procedure. The threshold of detection of two-dimensionally separated deoxyribonuclease I using the casein wash procedure was 1 picogram.
McGrew
B R
BR
Department of Biochemistry, University of New Hampshire, Durham 03824.
Green
D M
DM
D001345:Autoradiography; D002021:Buffers; D002364:Caseins; D002623:Chemistry Techniques, Analytical / Q000379:methods; D003902:Detergents / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D012710:Serum Albumin, Bovine / Q000494:pharmacology; D012967:Sodium Dodecyl Sulfate; D013462:Sulfur Radioisotopes; D001616:beta-Galactosidase / Q000378:metabolism
D002021:Buffers; D002364:Caseins; D003902:Detergents; D013462:Sulfur Radioisotopes; D012710:Serum Albumin, Bovine; D012967:Sodium Dodecyl Sulfate; D001616:beta-Galactosidase