Enumeration of rat allospecific antibody-producing cells by a solid-phase antibody-forming cell focus (AFCF) assay.
Individual rat cells producing antibody to rat MHC class I alloantigens ( RT1A ) and other erythrocyte (E)-associated alloantigens can be enumerated by a solid-phase antibody-forming cell focus ( AFCF ) assay. Cells are incubated in solid agarose medium in petri dishes coated with affinity-purified anti-immunoglobulin antibodies and antibody produced by any cell is captured by these anti-immunoglobulin antibodies. Foci of antibody to E alloantigens produced by single cells become visible upon subsequent addition of the appropriate target E. Execution of the assay in solid agarose medium facilitates handling of the petri dishes and reduces target background binding compared to performance of the assay in liquid medium.
D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000502:physiology; D000921:Antibody-Producing Cells / Q000276:immunology* / Q000378:metabolism; D001666:Binding Sites, Antibody; D001789:Blood Group Antigens / Q000276:immunology*; D002452:Cell Count / Q000379:methods; D000939:Epitopes; D006649:Histocompatibility Antigens / Q000276:immunology; D007118:Immunoassay / Q000379:methods*; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000378:metabolism; D007075:Immunoglobulin M / Q000096:biosynthesis / Q000378:metabolism; D007518:Isoantibodies / Q000096:biosynthesis; D051381:Rats; D011914:Rats, Inbred BN; D011917:Rats, Inbred Lew; D013154:Spleen / Q000166:cytology
D000888:Antibodies, Anti-Idiotypic; D001789:Blood Group Antigens; D000939:Epitopes; D006649:Histocompatibility Antigens; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D007518:Isoantibodies; C038353:histocompatibility antigens RT, rat