Use of a Western blotting technique in the purification of a cysteine proteinase inhibitor

✍ Scribed by Marcus J. Korth; Donald J. Finn; Gary L. Gustafson

• Publisher: Elsevier Science
• Year: 1988
• Tongue: English
• Weight: 297 KB
• Volume: 169
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(88)90270-9

✦ Synopsis

In Western blotting procedures, proteins are resolved in sodium dodecyl sulfate-polyacrylamide gels with subsequent electrophoretic transfer onto nitrocellulose membranes. Although this procedure is generally employed as an analytical technique for assessing interactions of proteins with antibodies, the present report describes the use of Western blotting as a preparative procedure in the purification of a biologically active proteinase inhibitor from the cellular slime mold, Dictyostelium discoideum. The feasibility of using Western blotting for inhibitor purification depended upon the unique stability properties of the inhibitor under denaturing conditions.