BASIC-PROTEINS AND BASIC MEMBRANES - ADJUSTING BLOTTING AND STAINING CONDITIONS TO IMMOBILON CD

被引：1

作者：

GIANAZZA, E ^{[1
]}

COARI, P ^{[1
]}

LOVATI, MR ^{[1
]}

MANZONI, C ^{[1
]}

GHIBAUDI, E ^{[1
]}

SALMONA, M ^{[1
]}

机构：

[1] IST RIC FARMACOL MARIO NEGRI, I-20157 MILAN, ITALY

来源：

JOURNAL OF CHROMATOGRAPHY A | 1995年 / 698卷 / 1-2期

关键词：

D O I：

10.1016/0021-9673(94)00761-W

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Transfer efficiency from polyacrylamide gels and binding to Immobilon P and CD were tested in different buffers with I-125-labelled proteins. With a derivatized poly(vinylidene difluoride) membrane, a pH 8 medium was found to be superior to a more alkaline solution, for both acidic and basic proteins. New staining protocols were tried on Immobilon CD. Toluidine Blue and iodine vapour gave a negative and a positive stain, respectively, with a fair band-to-background contrast. Protein sequencing after both stains was not impaired by interfering peaks. Biuret solution stained the protein bands pale pink but, even after copper removal with a chelating agent, it completely prevented Edman degradation. The first two procedures compare favourably with a commercial kit for protein detection, Quick Stain, that provides comparable sensitivity but results in several spurious peaks on protein sequencing.

引用

页码：351 / 359

页数：9

共 41 条

[1] CONTROLLED SYNTHESIS OF HBSAG IN A DIFFERENTIATED HUMAN-LIVER CARCINOMA-DERIVED CELL-LINE [J].