Immunoblot for αIIb of 293T cells transfected with mutant αIIbβ3 receptors.

Cells were solubilized in Triton X-100, electrophoresed into a 7% polyacrylamide gel under reduced conditions, and electrotransferred to PVDF membranes. Equal amounts of protein (150 μg), as determined by the BCA assay, were loaded per lane. The membrane was analyzed for αIIb using the mAb PMI-1. (A) Schematic diagram showing the processing and molecular weights of pro-αIIb in the endoplasmic reticulum and of mature αIIb in the Golgi. (B) Immunoblot of αIIb showing pro-αIIb and mature forms of control and mutant subunits.