Two-dimensional isoelectric focusing of P-selectin in platelets.

Triton X-100 lysates were made from resting untreated platelets (−DPDPB) or platelets treated with DPDPB (+DPDPB) and were subjected to electrophoresis on isoelectric focusing slab gels, as described in “Materials and methods.” The positions of IEF standards electrophoresed simultaneously are indicated. Gel lanes were then excised and subjected to electrophoresis in the second dimension on nonreducing 6% SDS-PAGE gels. Aliquots of appropriate lysate were run alongside in the reference well (REF.) for comparison. Samples were analyzed by immunoblotting using the P-selectin monoclonal antibody AC1.2.