Fig. 1. JAQ1, JAQ2, and JAQ3 bind to different epitopes on mouse GPVI. / (A) Whole platelet proteins were separated by SDS-PAGE under nonreducing conditions and immunoblotted with the indicated antibodies. Bound mAb was detected by HRP-labeled rabbit antirat Ig and ECL (upper, WB). Surface biotinylated platelets were lysed and immunoprecipitation was performed with the indicated antibodies. Precipitated proteins were detected with streptavidin-HRP and ECL (lower, IP). (B) Heparinized prp was incubated with JAQ1, JAQ2, JAQ3, or irrelevant IgG2a (all 20 μg/mL) followed by addition of polyclonal rabbit antirat immunoglobulin antibody (10 μg/mL) and light transmission was recorded on a Fibrintimer 4 channel aggregometer. (C) Platelets were preincubated with the indicated antibodies (20 μg/mL, 30 minutes) and washed; binding of FITC-labeled JAQ1, JAQ2, or JAQ3 was detected by flow cytometry.
Fig. 1.

JAQ1, JAQ2, and JAQ3 bind to different epitopes on mouse GPVI.

(A) Whole platelet proteins were separated by SDS-PAGE under nonreducing conditions and immunoblotted with the indicated antibodies. Bound mAb was detected by HRP-labeled rabbit antirat Ig and ECL (upper, WB). Surface biotinylated platelets were lysed and immunoprecipitation was performed with the indicated antibodies. Precipitated proteins were detected with streptavidin-HRP and ECL (lower, IP). (B) Heparinized prp was incubated with JAQ1, JAQ2, JAQ3, or irrelevant IgG2a (all 20 μg/mL) followed by addition of polyclonal rabbit antirat immunoglobulin antibody (10 μg/mL) and light transmission was recorded on a Fibrintimer 4 channel aggregometer. (C) Platelets were preincubated with the indicated antibodies (20 μg/mL, 30 minutes) and washed; binding of FITC-labeled JAQ1, JAQ2, or JAQ3 was detected by flow cytometry.

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