BSA standards or protein samples (10 μg/mL) are adsorbed onto a 96-well plate for 2 hrs at 37ºC. The BPDE-protein adducts present in the sample or standard are probed with an Anti-BPDE-I Antibody, followed by an HRP Conjugated Secondary Antibody. The BPDE protein adduct content in an unknown sample is determined by comparing with a standard curve that is prepared from predetermined BPDE-BSA standards.
BPDE-BSA Standard Curve.