Isolation of H-2 Alloantigens Solubilized by the Detergent NP-40

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Abstract
Murine tumor cells of two different H-2 genotypes (H-2b and H-2d) were radiolabeled in suspension culture with 3H-fucose, and intact, labeled cells were extracted with the non-ionic detergent NP-40. Solubilized native molecules bearing H-2 alloantigenic activity were isolated from other cell constituents by indirect immuno-precipitation. SDS-polyacrylamide gel electrophoretic analysis of specific precipitates of 3H-fucose-labeled H-2 alloantigens resulted in radiolabel patterns, that gave single peaks for each mouse strain examined. These peaks were proved to be H-2 alloantigens by competition experiments in which the alloantiserum used in the indirect precipitation was pretreated with a papain-solubilized, unlabeled, highly purified splenic H-2 alloantigen preparation of the same H-2 type. The efficiency of the NP-40 solubilization procedure was estimated to be nearly 100%.

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