Crossed Immunoelectrophoresis for the Study of Platelet Membrane Proteins

  • Simon Karpatkin
Part of the Blood Cell Biochemistry book series (BLBI, volume 2)

Abstract

The technique of crossed immunoelectrophoresis (CIE) of platelet membranes was introduced for the study of platelet membrane cell surface antigens (Hagen et al., 1979; Shulman and Karpatkin, 1980) because of several advantages that it has over sodium dodecyl sulfate—polyacrylamide gel electrophoresis (SDS—PAGE): (1) it does not completely denature membrane proteins, and thus intrinsic biologic activity can often be assayed (Kunicki et al., 1981; Howard et al., 1982; Hagen et al., 1981, 1982; Gogstad et al., 1982a,b, 1983; Gogstad and Brosstad, 1983; Karpatkin et al., 1984); (2) it is 10 times more sensitive than SDS—PAGE stained with Coomassie blue (Shulman and Karpatkin, 1980); (3) it may be used quantitatively, since the peak areas of individual immunoprecipitate arcs are proportional to the antigen/antibody ratios (Karpatkin et al., 1984); (4) various lectins, antibodies, or other ligands can be employed in intermediate spacer gels, providing an immunoaffinoelectrophoresis pattern (Hagen et al., 1979, 1980; Shulman and Karpatkin, 1980; Varon and Karpatkin, 1983); (5) conclusions can be drawn regarding the relative surface location of various membrane antigens by comparative CIE studies of antimembrane antiserum absorbed with whole cells as well as isotopic labeling of the intact platelet surface (Hagen et al., 1979; Shulman and Karpatkin, 1980); (6) amphiphilic proteins can be recognized by charge-shift CIE and crossed hydrophobic-interaction immunoelectrophoresis (Hagen et al., 1979) (examples of these advantages will be detailed below); and (7) charged antigenic components can be identified by mobility shifts of the antigen following incubation of the antigen with various enzymes, prior to electrophoresis (Karpatkin et al., 1986).

Keywords

Human Platelet Factor Xiii Platelet Membrane Ionic Detergent Major Antigen 
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Copyright information

© Springer Science+Business Media New York 1991

Authors and Affiliations

  • Simon Karpatkin
    • 1
  1. 1.New York University Medical SchoolNew YorkUSA

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