Jump to Main Content
Analysis of immune response: comparison of immunoblots after isoelectric focusing and sodium dodecyl sulfateâpolyacrylamide gel electrophoresis using cytoplasmic protein extract from Brucella
- Zygmunt, Michel S., Martin, JeanâCharles, Dubray, GÃ©rard
- FEMS microbiology letters 1990 v.70 no.3 pp. 263-268
- Brucella melitensis, antigens, bacteria, goats, immune response, immunoblotting, isoelectric focusing, pH, polyacrylamide gel electrophoresis, proteins, silver
- Analysis of the immune response towards the facultative intracellular bacterium, Brucella melitensis, was studied by immunoblotting after either isoelectric focusing (IEF) or sodium dodecyl sulfateâpolyacrylamide gel electrophoresis (SDSâPAGE). 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 4.5â5.4. CPE analysed by SDSâPAGE showed more than 20 silver stained proteins in the molecular range of 16â18 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 SDSâPAGE treatment, this technique may be best suited for analysis of the overall response to natural infection.