Characterization of the antihuman cellubrevin antibody.

(A) Gel-filtered platelets (50 μL) were permeabilized with 4 U/mL SL-O in the presence of reducing buffer (no addition) or 2 μM tetanus toxin in reducing buffer (tetanus toxin) for 45 minutes. Platelet proteins were subsequently solubilized in sample buffer, separated by SDS-PAGE, and transferred to a polyvinylidene (PVDF) membrane. Human cellubrevin was visualized by immunoblotting using the antihuman cellubrevin antibody. (B) Proteins from bovine brain, human fibroblasts, and human platelets were solubilized, separated by SDS-PAGE on 15% acrylamide gels, and transferred to PVDF membranes. Lysates were equalized for total protein, and immunoblotting was performed using either the antihuman cellubrevin antibody or an antibody directed against an epitope common to VAMP 1, VAMP 2, and human cellubrevin.