2-DIMENSIONAL MAPS IN VERY ACIDIC IMMOBILIZED PH GRADIENTS

被引：14

作者：

SINHA, PK

KOTTGEN, E

STOFFLERMEILICKE, M

GIANAZZA, E

RIGHETTI, PG

机构：

[1] UNIV MILAN,FAC PHARM,VIA CELORIA 2,I-20133 MILAN,ITALY

[2] UNIV MILAN,DEPT BIOMED SCI & TECHNOL,I-20133 MILAN,ITALY

[3] FREE UNIV BERLIN,KLINIKUM RUDOLF VIRCHOW,INST CLIN CHEM & BIOCHEM,BERLIN,GERMANY

[4] FREE UNIV BERLIN,DEPT VIROL,BERLIN,GERMANY

来源：

JOURNAL OF BIOCHEMICAL AND BIOPHYSICAL METHODS | 1990年 / 20卷 / 04期

关键词：

Immobilized pH gradient; Isoelectric focusing; Two-dimensional map;

D O I：

10.1016/0165-022X(90)90096-U

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Up to the present time it has been impossible to perform two-dimensional (2-D) separations in very acidic immobilized pH gradients (IPG), due to the lack of suitable buffering acrylamido derivatives to be incorporated into the polyacrylamide matrix. The advent of the pK 3.1 buffer (2-acrylamido glycolic acid; Righetti et al., J. Biochem. Biophys. Methods 16, 1988, 185-192) allowed the formulation of such acidic gradients. We report here separations in IPG pH 2.8-5.0 intervals of polypeptide chains from total lysates of rat intestinal and liver cells and 30S and 50S ribosomal proteins from Halobacterium marismortui. Conditions are given for highly reproducible first and second dimensions gels and for a proper silver staining of 2-D maps with practically no background deposition. © 1990.

引用

页码：345 / 352

页数：8

共 22 条

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