American Society of Hematology

Fig. 1.

$Fig. 1. In vitro phosphorylation of PI3K from resting or thrombin-stimulated platelets. Control (CO) or thrombin-treated (THR) platelets were lysed in 1% Triton-containing lysis buffer and Triton-soluble fractions were immunoprecipitated with equal amounts of either p85 antibody (lanes 2 and 4) or an isotype-matched control antibody (lanes 1 and 3). The washed immune complexes were incubated in the presence of γ-32P-ATP and Mn2+ in an in vitro kinase assay as described in Materials and Methods. Phosphorylated proteins were eluted, separated by SDS-PAGE and detected by autoradiography. Molecular mass markers in kilodaltons are indicated at the left.$

In vitro phosphorylation of PI3K from resting or thrombin-stimulated platelets. Control (CO) or thrombin-treated (THR) platelets were lysed in 1% Triton-containing lysis buffer and Triton-soluble fractions were immunoprecipitated with equal amounts of either p85 antibody (lanes 2 and 4) or an isotype-matched control antibody (lanes 1 and 3). The washed immune complexes were incubated in the presence of γ-³²P-ATP and Mn²⁺ in an in vitro kinase assay as described in Materials and Methods. Phosphorylated proteins were eluted, separated by SDS-PAGE and detected by autoradiography. Molecular mass markers in kilodaltons are indicated at the left.

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