Open AccessAnalysis of immune response: comparison of immunoblots after isoelectric focusing and sodium dodecyl sulfate‐polyacrylamide gel electrophoresis using cytoplasmic protein extract from Brucella
Author(s) -
Zygmunt Michel S.,
Martin JeanCharles,
Dubray Gérard
Publication year - 1990
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1990.tb13986.x
Subject(s) - isoelectric focusing , polyacrylamide gel electrophoresis , gel electrophoresis , sodium dodecyl sulfate , brucella , biology , microbiology and biotechnology , brucella melitensis , brucellaceae , chemistry , biochemistry , virology , enzyme , brucellosis
1. Summary Analysis of the immune response towards the facultative intracellular bacterium, Brucella melitensis , was studied by immunoblotting after either isoelectric focusing (IEF) or sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE). A cytoplasmic extract (CPE) of Brucella melitensis was used as antigen to analyse the response in 17 sera from naturally infected goats. CPE analysed by IEF exhibited 25 proteins within the pH range of 4.35 to 6. Immunoblotting revealed most of the stained bands around pH 4.5‐5.4. CPE analysed by SDS‐PAGE showed more than 20 silver stained proteins in the molecular range of 16–18 kDa to 70 kDa but immunoblotting revealed only 1 to 6 bands according to the sera tested. Because proteins are preserved in their native state with IEF, in contrast to SDS‐PAGE treatment, this technique may be best suited for analysis of the overall response to natural infection.