Isolation of Single-Stranded DNA Using Denaturing DNA Chromatography

✍ Scribed by Mark Dickman; David P. Hornby

• Publisher: Elsevier Science
• Year: 2000
• Tongue: English
• Weight: 58 KB
• Volume: 284
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1006/abio.2000.4669

✦ Synopsis

The versatility of the modified micro-Bradford CaPE assay was supported by the results with the strongly glycosylated glycophorin, which did not deviate excessively from the other protein preparations either in the rate of color development or in the sensitivity, whereas it showed very slow color development in the micro-Bradford assay (10) and extremely low absorbance in the Lowry DOC/TCA method.

Summary. Our modified micro-Bradford CaPE assay provided reliable results for BSA, purified membrane proteins, and membrane vesicle proteins. Interfering substances (detergents, lipids, and sulfhydryl reagents) were efficiently removed by washings with 70% ethanol. The disadvantages are the limited rectilinear range of the calibration curves for membrane proteins and the relatively large variation among the sensitivities for different membrane proteins. This fast, sensitive, robust, and inexpensive assay shows potential for widespread use.