Elution of proteins from sodium dodecyl sulfate-polyacrylamide gels, removal of sodium dodecyl sulfate, and renaturation of enzymatic activity: Results with sigma subunit of Escherichia coli RNA polymerase, wheat germ DNA topoisomerase, and other enzymes
✍ Scribed by Dayle A. Hager; Richard R. Burgess
- Publisher
- Elsevier Science
- Year
- 1980
- Tongue
- English
- Weight
- 965 KB
- Volume
- 109
- Category
- Article
- ISSN
- 0003-2697
No coin nor oath required. For personal study only.
✦ Synopsis
tained from Worthington (Freehold, N. J.). Wheat germ DNA topoisomerase was purified in our laboratory according to a procedure to be published elsewhere (W. Dynan. J. Jendrisak. D. Hager. and R. Burgess, unpublished procedure). Wheat cu-amylase was provided by R. Boston and plasmid pBR322 DNA was the gift of W. Dynan. Pentex five-times recrystallized bovine serum albumin (BSA) and poly(C) were purchased from Miles Laboratories, Inc. (Elkhart, Ind.), P-lactoglobulin, onitrophenyl-p-D-galactopyranoside and pnitrophenylphosphate were from Sigma Chemical Company (St. Louis, MO.). SDS, acrylamide, bisacrylamide, TEMED. ammonium persulfate, Coomassie brilliant blue R-250, Dowex AC l-X2, and Bio-Beads SM-2 were from BioRad laboratories (Richmond, Ca.). Guanidine-HCl was purchased from Schwarz/Mann (Orangeburg, N. Y.), urea from Mallinckrodt, Inc. (Paris, KY.). acetone (Gold Label) spectrophotometric grade from Aldrich Chemical Company, Inc. (Milwaukee, Wis.). and Spectru Cationic Blue from Pierce Chemical Company (Rockford.
Ill.). Soluene-100 was purchased from Packard (Downers Grove. Ill.). ATP, UTP. CTP. and GTP were obtained from P-L Biochemicals (Milwaukee, Wis.). [y-'"PIATP was prepared by the method of Schendel and Wells (7). ["H]UTP was purchased from Amersham (Arlington Heights, Ill.). T4 DNA was prepared as described by Gross et ~1. (5).
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