Analysis of serum antibody repertoires by isoelectric focusing and capillary blotting onto nitrocellulose paper.
A rapid method for the analysis of antigen-specific human serum antibodies is described. After isoelectric focusing on thin agarose gels, serum proteins are transferred to nitrocellulose paper by capillary blotting. The kinetics of transfer indicated that ca. 90% of 125I-labelled isolated human immunoglobulins (IgG, IgA and IgM) are transferred to the nitrocellulose paper within 10 min. Probing of the nitrocellulose blot with 125I-labelled tetanus toxoid antigen followed by autoradiography reveals the total serum anti-tetanus toxoid antibody repertoire, including high molecular weight IgM. This procedure is rapid (results can be obtained in less than 12 h), sensitive and should prove very useful for detailed studies and analysis of antibody repertoires in body fluids and extracts
A
A
Schibeci
A W
AW
Wade
W T
WT
Depew
M R
MR
Szewczuk
D000328:Adult; D000906:Antibodies / Q000032:analysis*; D001345:Autoradiography; D003101:Collodion; D004587:Electrophoresis, Agar Gel / Q000379:methods; D006801:Humans; D007106:Immune Sera / Q000032:analysis; D007075:Immunoglobulin M / Q000032:analysis; D007525:Isoelectric Focusing / Q000379:methods*; D010209:Paper; D013745:Tetanus Toxoid
D000906:Antibodies; D007106:Immune Sera; D007075:Immunoglobulin M; D013745:Tetanus Toxoid; D003101:Collodion