HUMAN MULTIPLE TISSUE WESTERN BLOTS - A NEW IMMUNOLOGICAL TOOL FOR THE ANALYSIS OF TISSUE-SPECIFIC PROTEIN EXPRESSION

Human multiple tissue Western (MTW) blots are premade immunoblots prep ared using proteins isolated from adult human tissue. The proteins are isolated from whole tissue homogenates under conditions designed to m inimize proteolysis and to ensure maximal representation of tissue-spe cific proteins. Sodium dodecyl sulfate (SDS) solubilized proteins are fractionated by SDS polyacrylamide gel electrophoresis and electroblot ted onto polyvinylidene fluoride membranes to generate blots ready for incubation with researcher supplied antibodies. Each lane of an MTW b lot contains an equivalent amount of total protein, allowing for the a nalysis of tissue-specific expression of a particular protein(s). The utility of MTW blots for Western blot applications was demonstrated by the detection of various cytoskeletal proteins and members of the ann exin family of calcium-dependent membrane-binding proteins. Several of these antigens were detected in separate cycles of antibody incubatio ns using the same MTW blot. This approach is possible using a strippin g procedure that allows the researcher to selectively remove both prim ary and secondary antibodies in a single incubation. The ease of multi ple reprobings makes MTW blots both economical and convenient research tools for western blot analyses of human tissue-specific proteins.