Multiple immunoblots after non-electrophoretic bidirectional transfer of a single SDS-PAGE gel with multiple antigens

被引:20
作者
Kurien, BT
Scofield, RH
机构
[1] UNIV OKLAHOMA, HLTH SCI CTR, DEPT MED, OKLAHOMA CITY, OK USA
[2] VET AFFAIRS MED CTR, OKLAHOMA CITY, OK 73104 USA
关键词
bidirectional transfer; SDS-PAGE; immunoblotting;
D O I
10.1016/S0022-1759(97)00052-5
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Western blotting is a very sensitive and powerful fundamental technique in immunology that has been used to detect and characterize proteins of low abundance. This technique employs the transfer of proteins separated on SDS-PAGE to nitrocellulose sheets for further detection using antibodies. Here we report the non-electrophoretic transfer of the 60-kDa Ro (or SSA) autoantigen, 240 and 220 kDa spectrin antigens and prestained molecular weight standards from SDS-PAGE gels to nitrocellulose to obtain multiple immunoblots. In fact, we have used this procedure to obtain 12 immunoblots from a single gel with multiple sera.
引用
收藏
页码:91 / 94
页数:4
相关论文
共 12 条
[1]   MOLECULAR ANALYSIS OF THE 60-KDA HUMAN RO RIBONUCLEOPROTEIN [J].
DEUTSCHER, SL ;
HARLEY, JB ;
KEENE, JD .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1988, 85 (24) :9479-9483
[2]   PRESENCE OF ANTI-LA(SS-B) IS ASSOCIATED WITH BINDING TO THE 13-KD CARBOXYL TERMINUS OF 60-KD RO(SS-A) IN SYSTEMIC LUPUS-ERYTHEMATOSUS [J].
DICKEY, WD ;
VANEGMOND, JE ;
HARDGRAVE, KL ;
HARLEY, JB ;
SCOFIELD, RH .
JOURNAL OF INVESTIGATIVE DERMATOLOGY, 1993, 100 (04) :412-416
[3]   PREPARATION AND CHEMICAL CHARACTERISTICS OF HEMOGLOBIN-FREE GHOSTS OF HUMAN ERYTHROCYTES [J].
DODGE, JT ;
HANAHAN, DJ ;
MITCHELL, C .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1963, 100 (01) :119-&
[4]   ELECTROPHORETIC TRANSFER OF PROTEINS FROM SODIUM DODECYL-SULFATE POLYACRYLAMIDE GELS TO A POSITIVELY CHARGED MEMBRANE-FILTER [J].
GERSHONI, JM ;
PALADE, GE .
ANALYTICAL BIOCHEMISTRY, 1982, 124 (02) :396-405
[5]   AFFINITY IMMUNOBLOTTING - HIGH-RESOLUTION ISOELECTRIC-FOCUSING ANALYSIS OF ANTIBODY CLONOTYPE DISTRIBUTION [J].
KNISLEY, KA ;
RODKEY, LS .
JOURNAL OF IMMUNOLOGICAL METHODS, 1986, 95 (01) :79-87
[6]   CLEAVAGE OF STRUCTURAL PROTEINS DURING ASSEMBLY OF HEAD OF BACTERIOPHAGE-T4 [J].
LAEMMLI, UK .
NATURE, 1970, 227 (5259) :680-+
[7]   VACUUM-BLOTTING - A NEW SIMPLE AND EFFICIENT TRANSFER OF PROTEINS FROM SODIUM DODECYL-SULFATE POLYACRYLAMIDE GELS TO NITROCELLULOSE [J].
PEFEROEN, M ;
HUYBRECHTS, R ;
DELOOF, A .
FEBS LETTERS, 1982, 145 (02) :369-372
[8]   PROTEIN TRANSFER FROM ISOELECTRIC-FOCUSING GELS - THE NATIVE BLOT [J].
REINHART, MP ;
MALAMUD, D .
ANALYTICAL BIOCHEMISTRY, 1982, 123 (02) :229-235
[9]   TRANSFER OF PROTEINS FROM GELS TO DIAZOBENZYLOXYMETHYL-PAPER AND DETECTION WITH ANTISERA - METHOD FOR STUDYING ANTIBODY SPECIFICITY AND ANTIGEN STRUCTURE [J].
RENART, J ;
REISER, J ;
STARK, GR .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1979, 76 (07) :3116-3120
[10]   THE BIDIRECTIONAL TRANSFER OF DNA AND RNA TO NITROCELLULOSE OR DIAZOBENZYLOXYMETHYL-PAPER [J].
SMITH, GE ;
SUMMERS, MD .
ANALYTICAL BIOCHEMISTRY, 1980, 109 (01) :123-129