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Use of 5′-UTP-agarose for ribonuclease affinity chromatography

✍ Scribed by Gary K. Smith; Keith J. Schray; Stephen W. Schaffer

Publisher: Elsevier Science
Year: 1978
Tongue: English
Weight: 460 KB
Volume: 84
Category: Article
ISSN: 0003-2697
DOI: 10.1016/0003-2697(78)90058-1

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✦ Synopsis

The use of commercially available 5'-UTP-agarose as an affinity chromatography resin for RNase has been described. It was shown that at pH 5.3, 0.025 M piperazine-HCI buffer was effective for the adsorption of active RNase A and exhibited little nonbiospecific binding as has been shown earlier for Sepharose-aPhpUp [Stewart, G. R., and Stevenson, K. J. (1973) Biochem. J. 135, 427-4411. Phosphate buffer at either pH 3.0 or 5.45 eluted essentially all of the RNase activity added to the column; however, pH 5.45 was slightly more efficient. Competitive elution experiments with 2'(3')-UMP yielded a linear plot of l/(V -V,) vs [I].

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