A DOT-BLOT ASSAY FOR QUANTITATION OF NANOGRAM AMOUNTS OF PROTEIN IN THE PRESENCE OF CARRIER AMPHOLYTES AND OTHER POSSIBLY INTERFERING SUBSTANCES

被引：33

作者：

GUTTENBERGER, M ^{[1
]}

NEUHOFF, V ^{[1
]}

HAMPP, R ^{[1
]}

机构：

[1] MAX PLANCK INST EXPTL MED,FORSCHUNGSSTELLE NEUROCHEM,W-3400 GOTTINGEN,GERMANY

来源：

ANALYTICAL BIOCHEMISTRY | 1991年 / 196卷 / 01期

关键词：

D O I：

10.1016/0003-2697(91)90124-C

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A method for protein determination in one- and two-dimensional electrophoresis sample buffer is presented. Accurate quantitation of protein in two-dimensional electrophoresis sample buffer (9.5 m urea, 2% Nonidet P-40, 2% carrier ampholytes, and 5% 2-mercaptoethanol) required removal of carrier ampholytes prior to the assay. This was made possible by taking advantage of the mutual solubility/insolubility of carrier ampholytes/proteins in saturated ammonium sulfate solution. In addition, improvement of protein determination in denaturing electrophoresis sample buffer containing the anionic detergent sodium dodecyl sulfate and the reducing agent 2-mercaptoethanol was achieved. The assay covers a range of sensitivity from 40 ng to 20 μg of protein. The procedure is applicable to large numbers of samples. © 1991.

引用

页码：99 / 103

页数：5

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