Immunoelectrophoretic analyses of antigenic and human specific proteins of red cell membrane.
Rabbit antisera were raised for red cell stroma extracted with each of the following solutions; 5 mM phosphate buffer pH 8.0 (A antigen). 5 mM EDTA containing 5 mM 2-mercaptoethanol (B), 1/15 M phosphate buffered saline (C), 0.05% SDS (D), 50 mM Tris-HCl buffer pH 8.0 containing 1% SDS, 1 mM EDTA and 1% 2-mercaptoethanol (E) and 1% Triton X-100 (F). Crossed immunoelectrophoresis showed that E antigen produced relatively numerous precipitation lines, 7-9, against each of the antisera, suggesting that E solution was superior to the others for membrane solubilization. However, the Ouchterlony test demonstrated potent precipitin activity in anti-C, and the antiserum absorbed with monkey stroma which was extracted with F solution was proved to have strict human specificity. By adopting the antiserum to affinity chromatography, human specific antigens were isolated. The antigens gave four bands moving faster than Band 5 at SDS-polyacrylamide gel electrophoresis none of which was stained by Schiff's reagent. Using absorbed anti-C, human specificity of blood stains kept us to for two years could be identified.
D000818:Animals; D000941:Antigens / Q000032:analysis*; D001801:Blood Stains; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000032:analysis*; D004910:Erythrocyte Membrane / Q000276:immunology* / Q000378:metabolism; D004912:Erythrocytes / Q000276:immunology*; D005554:Forensic Medicine; D000882:Haplorhini; D006801:Humans; D007123:Immunoelectrophoresis, Two-Dimensional; D008565:Membrane Proteins / Q000032:analysis / Q000276:immunology*; D011817:Rabbits
D000941:Antigens; D000939:Epitopes; D008565:Membrane Proteins