Membrane associated antigens of human malignant melanoma. I. Internal labeling, detergent solubilization and characterization by homologous antisera and polyacrylamide gel electrophoresis.
Cultured melanoma cells were labeled with 3H-leucine over a period of 1-3 days. The labeled cells were mechanically disrupted and a preparation of "extranuclear membranes" was obtained by differential centrifugation. The membrane fragments were solubilized by the nonionic detergent NP-40 and the soluble material was double precipitated with antisera from melanoma patients and anti-human immunoglobulin sera. Because of the small quantitative differences of precipitated radiolabeled material between control and melanoma patients' sera, the precipitates were further analyzed on SDS-containing polyacrylamide gels. The labeled profiles of experimental and control gels now revealed clearcut differences usually seen in 2-3 characteristic peaks in the molecular weight range from 130,000-330,000.
D000951:Antigens, Neoplasm / Q000032:analysis / Q000302:isolation & purification; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007106:Immune Sera; D008545:Melanoma / Q000276:immunology*; D008970:Molecular Weight; D012995:Solubility; D013347:Subcellular Fractions / Q000276:immunology; D013501:Surface-Active Agents
D000951:Antigens, Neoplasm; D007106:Immune Sera; D013501:Surface-Active Agents