Immunochemical reactivity of simian sarcoma virus polypeptides isolated by preparative sodium dodecyl sulfate polyacrylamide gel electrophoresis.
The polypeptides associated with a zonal centrifugation purified simian sarcoma virus propagated in lymphoblastoid NC-37 cells were isolated by preparative polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS) using a procedure designed to minimize the loss of immunochemical reactivity. The proteins p10, p15, p28, p36, p44, p75, and p86 were obtained in large yield and high degree of homogeneity. The electrophoretically purified p28 was analyzed by competition radioimmunoassay using antiserum to a pore exclusion and ion exchange purified simian sarcoma virus p28. Complete competition was observed with extracts of simian sarcoma virus infected cells. No competition was observed with uninfected or unrelated, infected cell extracts. The antigen-antibody affinity as measured by the slope of the competition curve using antiserum to p28 and 125I-labeled and electrophoretically purified p28 was the same as that for the p28 released from sonication-disrupted simian sarcoma virus. The data indicates that preparative purifications by polyacrylamide gel electrophoresis in the presence of SDS may be generally applicable for the isolation of proteins with essentially the same immunospecificities and affinity for a specific antiserum as proteins isolated by procedures that avoid the use of SDS and electrophoresis.
D000818:Animals; D000915:Antibody Affinity; D001358:Avian Sarcoma Viruses / Q000276:immunology; D002460:Cell Line; D002642:Chick Embryo; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008970:Molecular Weight; D010455:Peptides / Q000276:immunology* / Q000302:isolation & purification; D011863:Radioimmunoassay; D051381:Rats; D012190:Retroviridae / Q000276:immunology*; D012842:Sarcoma Virus, Woolly Monkey / Q000032:analysis / Q000276:immunology*; D012526:Satellite Viruses / Q000276:immunology; D014764:Viral Proteins / Q000276:immunology* / Q000302:isolation & purification
D010455:Peptides; D014764:Viral Proteins