Preparative polyacrylamide gel electrophoresis purification of Clostridium perfringens enterotoxin
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Preparative polyacrylamide gel electrophoresis purification of Clostridium perfringens enterotoxin.
Preparative polyacrylamide gel electrophoresis has been used to purify the enterotoxin of Clostridium perfringens from Sephadex G-100 extracts. Purified toxin of high specific activity was eluted in 1 to 3 h, depending upon the length of the acrylamide gel used. Recovery of biological activity with this technique ranged from 80 to 90%. The purity and physical characteristics of the toxin were s...
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Preparative polyacrylamide gel electrophoresis has been used in the final stages of purification to prepare horse serum cholinesterase which was at least 9.5% pure, as judged by analytical polyacrylamide gel electrophoresis. The starting material, already purified about 109-fold by the Strelitz method, was purified an additional 89-fold by the procedure. The method involved Sephadex G-ZOO and S...
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Anti-enterotoxin immunoglobulins immobilized on CH-Sepharose or CNBr-Sepharose were used for affinity chromatography purification of Clostridium perfringens enterotoxin. Cell extracts containing enterotoxin or partially purified toxin preparations were applied to the column and nonspecifically-bound protein was eluted. NaOH was used to elute specifically bound toxin. The purity of enterotoxin p...
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The sporulation-specific enterotoxin of Clostridium perfringens type A, which is the toxin active in human food poisoning, has been purified from extracts of sporulating cells. Highly purified enterotoxin was obtained by treatment of crude cell extract with ribonuclease for 30 min, followed by sequential chromatography on Sephadex G-100, Cellex T cellulose, and hydroxylapatite. Recovery was 65 ...
متن کاملProteolysis of Clostridium perfringens type A enterotoxin during purification.
The small satellite bands of enterotoxin frequently seen in polyacrylamide gels following purification of Clostridium perfringens enterotoxin were found to be due to endogenous protease activity and were not present if phenylmethylsulfonyl fluoride (PMSF; 1 mM) and EDTA (10 mM) were used in the purification protocol. The use of PMSF was avoided by passing gel filtration-purified enterotoxin mat...
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ژورنال
عنوان ژورنال: Infection and Immunity
سال: 1977
ISSN: 0019-9567,1098-5522
DOI: 10.1128/iai.17.2.425-429.1977