Figure 2.

Bacitracin, a thiol-isomerase inhibitor, inhibited αIIbβ3 activation of fibrinogen and PAC-1 binding. Fibrinogen and PAC-1 binding to activated human αIIbβ3 was inhibited by bacitracin. (A) Fibrinogen and bacitracin were added into a Mn²⁺-activated, purified, native human αIIbβ3 integrin precoated ELISA plate. After washing in PBS, anti-fibrinogen antibody was added, followed by HRP-conjugated secondary antibody. o-Phenylenediamine dihydrochloride (OPD) substrate was added and absorbance was measured at 492 nm. (B) PAC-1 and bacitracin were added into a Mn²⁺-activated, purified, native human αIIbβ3 integrin precoated ELISA plate. After washing in PBS, HRP-conjugated secondary antibody was added. OPD substrate was added and absorbance was measured at 492 nm. Mean ± SEM; *P < .05, ***P < .001, n = 4-6 each.