Inhibition of biotin–HKa binding to Mac-1 transfected cells by antibody and fibrinogen.

Biotin–HKa (10 nmol/L) was incubated with Mac-1 transfected or control transfected HEK 293 cells (4 × 10⁶/mL) in binding buffer containing 50 μmol/L ZnCl₂, 1 mmol/L MgCl₂, and 1 mmol/L CaCl₂ for 45 minutes at room temperature. The reactions were carried out either in the presence or absence of antibody, 2LPM19c, or fibrinogen as indicated. The relative amount of biotin–HKa bound to the Mac-1 transfected cells in the presence of inhibitors was determined by comparing that with biotin–HKa alone. Data are the mean ± SE of 3 separate experiments.