Expression of MoAb 62 LIBS epitope on A5 cells (A), VNRC3 cells (B), and resting platelets (C) by echistatin and its analogues. A5 or VNRC3 cells (5 × 105 of each) were incubated with echistatin (○), D27W echistatin (•), echistatin 1-41 (▿), or R24A echistatin (▾) for 15 minutes at room temperature. After being washed, MoAb 62 (1 μg per sample) was added for 45 minutes at 4°C. Cells were then incubated with FITC-conjugated goat antimouse IgG, fixed, and analyzed by flow cytometry as described in the Materials and Methods. In the case of platelets (6 × 106 per sample), the washing steps were omitted and incubations with antibodies were performed at room temperature for 30 minutes. Error bars represent the standard deviations for three independent experiments.
