Effect of beta-cyclodextrin on the renaturation of enzymes after sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
Activity gel assays require a long incubation time (several hours) on renaturation of enzymatic activity after sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). To reduce the incubation time, we used a novel renaturation buffer containing cyclic oligosaccharide beta-cyclodextrin (beta-CD) which is capable of capturing SDS. Yeast alpha-glucosidase, used as a model protein, was run on SDS-PAGE, and then the gel matrix was incubated in a variety of renaturation buffers. Compared with conventional renaturation buffers containing Triton X-100 or isopropanol, our novel renaturation buffer containing beta-CD can restore enzymatic activity within 10 min. Therefore, this new format represents a good alternative with reduced incubation time for activity gel assays.
Yamamoto
Etsushi
E
Department of Chemistry and Biotechnology, Graduate School of Engineering, University of Tokyo, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
Yamaguchi
Satoshi
S
Nagamune
Teruyuki
T
D004591:Electrophoresis, Polyacrylamide Gel; D004798:Enzymes / Q000737:chemistry*; D020673:Protein Renaturation; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry; D013194:Staining and Labeling; D000520:alpha-Glucosidases / Q000737:chemistry; D047392:beta-Cyclodextrins / Q000737:chemistry*
D004798:Enzymes; D047392:beta-Cyclodextrins; D012967:Sodium Dodecyl Sulfate; D000520:alpha-Glucosidases