Characterization of immune complex components by dot blot analysis.
A method is described for the characterization of immune complex components by dot blot analysis. After isolation by chromatographic techniques and precipitation with polyethylene glycol, immune complexes were dissociated in 0.1 M phosphate (pH 2) and bound to a nitrocellulose membrane in a dot blot unit. Biotinylated probes were then used to identify the following immune complex components: specific antigens, biologically active antibodies, antibody isotypes, antibody subclasses, antibody idiotypes, and rheumatoid factors. This nonradioactive procedure takes less than 2 h to perform and has been used to analyze immune complexes isolated from sera (rabbit and human) and synovial fluid (human).
Hoffman
W L
WL
Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas 75235.
Kelly
P J
PJ
Ruggles
A O
AO
D000143:Acids; D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000032:analysis; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex / Q000032:analysis* / Q000302:isolation & purification; D001172:Arthritis, Rheumatoid / Q000097:blood / Q000276:immunology; D002021:Buffers; D011232:Chemical Precipitation; D003101:Collodion; D015922:Complement C1q; D004586:Electrophoresis / Q000379:methods; D006801:Humans; D006863:Hydrogen-Ion Concentration; D015151:Immunoblotting / Q000379:methods*; D007074:Immunoglobulin G / Q000032:analysis; D007132:Immunoglobulin Isotypes / Q000032:analysis; D010047:Ovalbumin / Q000032:analysis; D011817:Rabbits; D013582:Synovial Fluid / Q000276:immunology
D000143:Acids; D000888:Antibodies, Anti-Idiotypic; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D002021:Buffers; D007074:Immunoglobulin G; D007132:Immunoglobulin Isotypes; D015922:Complement C1q; D003101:Collodion; D010047:Ovalbumin