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Use of avidin-sepharose to lsolate and identify biotin polypeptides from crude extracts

✍ Scribed by Judith A. Swack; Gary L. Zander; Merton F. Utter


Publisher
Elsevier Science
Year
1978
Tongue
English
Weight
833 KB
Volume
87
Category
Article
ISSN
0003-2697

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✦ Synopsis


A method of isolating and identifying biotin polypeptides from crude cellular extracts is described. Protein samples are run on small avidin-Sepharose columns. After washing away nonspecifically bound protein, the biotin enzymes are eluted using an SDS-urea solution. The inactive polypeptides are then electrophoresed on SDS-polyacrylamide gels. The biotin polypeptides in the gels are identified by using fluorescent avidin or by analyzing for radioactive biotin-labeled polypeptides. A sensitive method for assaying biotin using avidin-Sepharose is also described.