American Society of Hematology

Figure 1.

$Figure 1. Coimmunoprecipitation of CD151 with integrin αIIbβ3 from human and murine platelet lysates. (A) Platelet lysates (1.5 mg) solubilized with indicated detergents were immunoprecipitated with normal IgG1, integrin complex–specific αIIbβ3 Ab, P2, PECAM-1.2, and 11B1 (anti-CD151). Samples were resolved on 12.5% SDS-PAGE under nonreducing conditions and Western blotted for CD151 with 11B1 Ab. (B) Wild-type mouse platelets (150 × 109/L) were chemically cross-linked with 2 mM DTSSP and then solubilized with 1% Triton X-100 and Triton-soluble fractions immunoprecipitated with isotype control, CD9, integrin αIIbβ3 complex–specific mAb, and 11B1 (CD151) (left). Murine and human platelet lysates (100 μg) blotted with 11B1 Ab are included as positive markers of CD151 recognition (right). Samples were resolved on 12.5% SDS-PAGE under nonreducing conditions and Western blotted for CD151 with biotinylated 11B1 Ab followed by horseradish peroxidase (HRP)–streptavidin. Glycos indicates glycosylated.$

Coimmunoprecipitation of CD151 with integrin α_IIbβ₃ from human and murine platelet lysates. (A) Platelet lysates (1.5 mg) solubilized with indicated detergents were immunoprecipitated with normal IgG₁, integrin complex–specific α_IIbβ₃ Ab, P2, PECAM-1.2, and 11B1 (anti-CD151). Samples were resolved on 12.5% SDS-PAGE under nonreducing conditions and Western blotted for CD151 with 11B1 Ab. (B) Wild-type mouse platelets (150 × 10⁹/L) were chemically cross-linked with 2 mM DTSSP and then solubilized with 1% Triton X-100 and Triton-soluble fractions immunoprecipitated with isotype control, CD9, integrin α_IIbβ₃ complex–specific mAb, and 11B1 (CD151) (left). Murine and human platelet lysates (100 μg) blotted with 11B1 Ab are included as positive markers of CD151 recognition (right). Samples were resolved on 12.5% SDS-PAGE under nonreducing conditions and Western blotted for CD151 with biotinylated 11B1 Ab followed by horseradish peroxidase (HRP)–streptavidin. Glycos indicates glycosylated.

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