A NONLINEAR WIDE-RANGE IMMOBILIZED PH GRADIENT FOR 2-DIMENSIONAL ELECTROPHORESIS AND ITS DEFINITION IN A RELEVANT PH SCALE

被引：344

作者：

BJELLQVIST, B ^{[1
]}

PASQUALI, C ^{[1
]}

RAVIER, F ^{[1
]}

SANCHEZ, JC ^{[1
]}

HOCHSTRASSER, D ^{[1
]}

机构：

[1] UNIV HOSP GENEVA,DEPT MED,CH-1211 GENEVA 4,SWITZERLAND

来源：

ELECTROPHORESIS | 1993年 / 14卷 / 12期

关键词：

D O I：

10.1002/elps.11501401209

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A new nonlinear immobilized pH gradient (IPG) is proposed as the first dimension for two-dimensional electrophoresis. In comparison to conventional carrier ampholyte techniques, it offers better resolution and greater reproducibility whilst allowing application of higher protein loads. Furthermore, we have checked and supplemented existing data on pK values for the immobilized groups in the presence of 8M urea. This is necessary for pH gradients to be defined in a pH scale relevant to the focusing conditions such that spot positions can be related to amino acid compositions. The data will allow definition of pH scales for the temperature range 10-25 degrees C and for a pH range covering the major part of the nonlinear pH gradient. With the latter, focusing positions are neither influenced by urea concentration nor by the choice or the concentration of detergent or carrier ampholyte. Temperature is the only parameter affecting focusing reproducibility and here any changes in focusing positions can be related to the amino acid compositions of peptides.

引用

页码：1357 / 1365

页数：9

共 21 条

[1] IPGMAKER - A PROGRAM FOR IBM-COMPATIBLE PERSONAL COMPUTERS TO CREATE AND TEST RECIPES FOR IMMOBILIZED PH GRADIENTS [J].

ALTLAND, K .

ELECTROPHORESIS, 1990, 11 (02) :140-147

[2] ISOELECTRIC-FOCUSING IN IMMOBILIZED PH GRADIENTS - PRINCIPLE, METHODOLOGY AND SOME APPLICATIONS [J].

BJELLQVIST, B ;

EK, K ;

RIGHETTI, PG ;

GIANAZZA, E ;

GORG, A ;

WESTERMEIER, R ;

POSTEL, W .

JOURNAL OF BIOCHEMICAL AND BIOPHYSICAL METHODS, 1982, 6 (04) :317-339

[3] THE FOCUSING POSITIONS OF POLYPEPTIDES IN IMMOBILIZED PH GRADIENTS CAN BE PREDICTED FROM THEIR AMINO-ACID-SEQUENCES [J].

BJELLQVIST, B ;

HUGHES, GJ ;

PASQUALI, C ;

PAQUET, N ;

RAVIER, F ;

SANCHEZ, JC ;

FRUTIGER, S ;

HOCHSTRASSER, D .

ELECTROPHORESIS, 1993, 14 (10) :1023-1031

[4] MICROPREPARATIVE 2-DIMENSIONAL ELECTROPHORESIS ALLOWING THE SEPARATION OF SAMPLES CONTAINING MILLIGRAM AMOUNTS OF PROTEINS [J].

BJELLQVIST, B ;

SANCHEZ, JC ;

PASQUALI, C ;

RAVIER, F ;

PAQUET, N ;

FRUTIGER, S ;

HUGHES, GJ ;

HOCHSTRASSER, D .

ELECTROPHORESIS, 1993, 14 (12) :1375-1378

[5] ISOELECTRIC POINTS OF PROTEINS, DETERMINED BY ISOELECTRIC-FOCUSING IN THE PRESENCE OF UREA AND ETHANOL [J].

GELSEMA, WJ ;

DELIGNY, CL ;

VANDERVEEN, NG .

JOURNAL OF CHROMATOGRAPHY, 1979, 171 (APR) :171-181

[6] NON-LINEAR PH COURSES WITH IMMOBILIZED PH GRADIENTS [J].

GIANAZZA, E ;

GIACON, P ;

SAHLIN, B ;

RIGHETTI, PG .

ELECTROPHORESIS, 1985, 6 (01) :53-56

[7] ON THE REPRODUCIBILITY OF BAND POSITION IN ELECTROPHORETIC SEPARATIONS [J].

GIANAZZA, E ;

ASTRUATESTORI, S ;

CACCIA, P ;

GIACON, P ;

QUAGLIA, L ;

RIGHETTI, PG .

ELECTROPHORESIS, 1986, 7 (02) :76-83

[8] ISOELECTRIC-FOCUSING IN IMMOBILIZED PH GRADIENTS IN PRESENCE OF UREA AND NEUTRAL DETERGENTS [J].

GIANAZZA, E ;

ARTONI, G ;

RIGHETTI, PG .

ELECTROPHORESIS, 1983, 4 (05) :321-326

[9] THE CURRENT STATE OF TWO-DIMENSIONAL ELECTROPHORESIS WITH IMMOBILIZED PH GRADIENTS [J].

GORG, A ;

POSTEL, W ;

GUNTHER, S .

ELECTROPHORESIS, 1988, 9 (09) :531-546

[10] ELIMINATION OF POINT STREAKING ON SILVER STAINED TWO-DIMENSIONAL GELS BY ADDITION OF IODOACETAMIDE TO THE EQUILIBRATION BUFFER [J].

GORG, A ;

POSTEL, W ;

WESER, J ;

GUNTHER, S ;

STRAHLER, JR ;

HANASH, SM ;

SOMERLOT, L .

ELECTROPHORESIS, 1987, 8 (02) :122-124

← 1 2 3 →