Affinity immunoblotting

Autor:	Keith A. Knisley, L. Scott Rodkey
Rok vydání:	1986
Předmět:	biology Chemistry Isoelectric focusing Microgram Immunology Molecular biology chemistry.chemical_compound Immune system Membrane Antigen Polyclonal antibodies biology.protein Immunology and Allergy Antibody Nitrocellulose
Zdroj:	Journal of Immunological Methods. 95:79-87
ISSN:	0022-1759
DOI:	10.1016/0022-1759(86)90320-0
Popis:	A sensitive and specific method is proposed for the analysis of specific antibody clonotype changes occurring during an immune response and for comparing multiple sera for antibody clonotype similarities. Polyclonal serum antibodies separated by isoelectric focusing (IEF) were analyzed by an affinity immunoblotting method using antigen-coated nitrocellulose membranes. Antibodies present on the surface of the acrylamide gels following IEF bind the antigen on the nitrocellulose when the coated nitrocellulose is laid over the gels. The technique has been used to analyze Ig clonotypes specific for five protein antigens and two carbohydrate antigens. Optimal antigen concentrations for coating the nitrocellulose membranes were found to range from 10-100 microgram/ml.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_________::fa59f2466b586bc2afa350df5683b4ba https://doi.org/10.1016/0022-1759(86)90320-0 Zobrazit plný text záznamu