THE USE OF SILVER STAIN IN SODIUM DODECYL-SULFATE POLYACRYLAMIDE-GEL ELECTROPHORESIS OF IMMUNOPRECIPITATES OBTAINED BY CROSSED IMMUNOELECTROPHORESIS AND LIQUID PRECIPITATION

被引:7
作者
CUTTS, JL
SWEET, GH
TEGTMEIER, GE
机构
[1] WICHITA STATE UNIV, DEPT BIOL SCI, WICHITA, KS 67208 USA
[2] COMMUNITY BLOOD CTR, KANSAS CITY, MO USA
关键词
D O I
10.1002/elps.1150050605
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Ag staining was examined as a means for detecting antigenic polypeptides separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) after recovery from immunoprecipitates produced by crossed immunoelectrophoresis and liquid precipitation. Both precipitin procedures resulted in detection of the expected polypeptides with bovine serum albumin (BSA) and human transferrin as model antigens, as well as microheterogeneity of BSA: antigenic polypeptides were rarely detected with Coomassie Brilliant Blue staining of SDS-PAGE. The technique is advantageous over the use of radiolabeled antigens in being less expensive and time consuming and over antibody-blot procedures in detecting antigenic activity prior to the denaturing conditions of SDS-PAGE. Possible draw backs are the appearance of antibody polypeptide chains on the electrophorograms as well as other non-antigenic components when crude extracts of virally-infected cells are used as antigen.
引用
收藏
页码:349 / 353
页数:5
相关论文
共 18 条
[1]   PROTEINS OF MURINE CYTOMEGALOVIRUS - IDENTIFICATION OF STRUCTURAL AND NONSTRUCTURAL ANTIGENS IN INFECTED-CELLS [J].
CHANTLER, JK ;
HUDSON, JB .
VIROLOGY, 1978, 86 (01) :22-36
[2]  
CROWLE AJ, 1973, IMMUNODIFFUSION, P22
[3]  
EMMETT M, 1982, J IMMUNOL METHODS, V50, pR65
[4]  
Foster J.F., 1977, ALBUMIN STRUCTURE FU, P53, DOI DOI 10.1016/B978-0-08-019603-9.50010-7
[5]  
HABEEB AFS, 1978, IMMUNOCHEMISTRY PROT, V1, P163
[6]   TANDEM-CROSSED IMMUNOELECTROPHORESIS [J].
KROLL, J .
SCANDINAVIAN JOURNAL OF IMMUNOLOGY, 1973, 2 :57-59
[7]   CLEAVAGE OF STRUCTURAL PROTEINS DURING ASSEMBLY OF HEAD OF BACTERIOPHAGE-T4 [J].
LAEMMLI, UK .
NATURE, 1970, 227 (5259) :680-+
[8]  
LOWRY OH, 1951, J BIOL CHEM, V193, P265
[9]   HUMAN CYTOMEGALOVIRUS-INDUCED IMMEDIATE EARLY ANTIGENS - ANALYSIS IN SODIUM DODECYL SULFATE-POLYACRYLAMIDE GEL-ELECTROPHORESIS AFTER IMMUNOPRECIPITATION [J].
MICHELSON, S ;
HORODNICEANU, F ;
KRESS, M ;
TARDYPANIT, M .
JOURNAL OF VIROLOGY, 1979, 32 (01) :259-267
[10]   POLYACRYLAMIDE-GEL ELECTROPHORETIC ANALYSIS OF HERPES-SIMPLEX VIRUS TYPE-1 IMMUNO-PRECIPITATES OBTAINED BY QUANTITATIVE IMMUNOELECTROPHORESIS IN ANTIBODY-CONTAINING AGAROSE-GEL [J].
NORRILD, B ;
VESTERGAARD, BF .
JOURNAL OF VIROLOGY, 1977, 22 (01) :113-117