Anti-human Integrin αIIbβ3 Recombinant Antibody (c3C7) (TAB-753LC) (CAT#: TAB-753LC)

Figure 1 The specific binding of c3C7 to CHO/αIIbβ3 cells.

CHO/αIIbβ3 cells grown in the 96-well plate. After fixing and blocking, the cells were incubated with supernatant containing 1 μg/mL c3C7 and 1 μg/mL human IgG. The supernatant of the CHO cells was added as the negative control. After washing, HRP conjugated goat anti-human IgG was used for detection and TMB substrate was used for color development. The OD of each well was read at 450 nm.

Jiang, A., Zhang, W., Wu, Q., Jin, W., Tang, Y., Zhang, J., & Liu, J. N. (2014). Construction and characterization of a novel chimeric antibody c3C7 specific for the integrin αIIbβ3 complex. Applied microbiology and biotechnology, 98(1), 105-114.

Figure 2 The specific binding of c3C7 to CHO/αIIbβ3 cells.

(A)CHO cells were washed and incubated with 1 μg/mL c3C7 followed by FITC-conjugated goat antihuman IgG. Then, the cells were washed and analyzed by flow cytometry. The black peak is the negative control. (B)CHO/αIIbβ3 cells were washed and incubated with 1 μg/mL c3C7 followed by FITC-conjugated goat antihuman IgG. Then, the cells were washed and analyzed by flow cytometry. The black peak is the negative control.

Jiang, A., Zhang, W., Wu, Q., Jin, W., Tang, Y., Zhang, J., & Liu, J. N. (2014). Construction and characterization of a novel chimeric antibody c3C7 specific for the integrin αIIbβ3 complex. Applied microbiology and biotechnology, 98(1), 105-114.

Figure 3 The reactivity of c3C7 with the integrin αIIbβ3 complex.

CHO/αIIbβ3 cells were pretreated with 5 mmol/L EDTA at 37 °C for 30 min and then incubated with 1 μg/mL c3C7 followed by FITC-conjugated goat antihuman IgG (gray peak). CHO/αIIbβ3 cells were not pretreated with 5 mmol/L EDTA and directly incubated with 1 μg/mL c3C7 followed by FITC-conjugated goat anti-human IgG (white peak). Cells stained directly with FITC-conjugated goat anti-human IgG acted as the negative control (black peak)

Jiang, A., Zhang, W., Wu, Q., Jin, W., Tang, Y., Zhang, J., & Liu, J. N. (2014). Construction and characterization of a novel chimeric antibody c3C7 specific for the integrin αIIbβ3 complex. Applied microbiology and biotechnology, 98(1), 105-114.

Figure 4 Specific reactivity of c3C7 with platelets of different animals.

Platelets were washed and incubated with 1 μg/mL c3C7 followed by FITCconjugated goat anti-human IgG. Then, the cells were washed and analyzed by flow cytometry. The black peak is the negative control.

Jiang, A., Zhang, W., Wu, Q., Jin, W., Tang, Y., Zhang, J., & Liu, J. N. (2014). Construction and characterization of a novel chimeric antibody c3C7 specific for the integrin αIIbβ3 complex. Applied microbiology and biotechnology, 98(1), 105-114.

Figure 5 The binding curve of c3C7 and 3C7 to the lysate CHO/αIIbβ3.

Microplates were coated with diluted lysate (4, 2, and 1 μg/mL) of CHO/αIIbβ3. After blocking, the plates were incubated with different concentrations (0.05, 0.075, 0.1, 0.25, 0.5, 0.75, 1, 1.5, 2, 5, 10, 12.5, 15, 18, and 20 nmol/L) of c3C7 (top panel) or 3C7 (bottom panel). After washing, HRP-conjugated goat anti-human IgG was used to detect c3C7 and HRP-conjugated goat anti-mouse IgG was used to detect 3C7. TMB substrate was used for color development. The OD of each well was read at 450 nm

Jiang, A., Zhang, W., Wu, Q., Jin, W., Tang, Y., Zhang, J., & Liu, J. N. (2014). Construction and characterization of a novel chimeric antibody c3C7 specific for the integrin αIIbβ3 complex. Applied microbiology and biotechnology, 98(1), 105-114.

Figure 6 The effect of c3C7 on human platelet aggregation.

Human PRP was incubated at various concentrations (0, 1, 2.5, 5, 10, 20, and 40 μg/mL) of c3C7 and 3C7 at 37 °C for 5 min. Platelet aggregation was induced by 10 μmol/L ADP. The maximal platelet aggregation within 5 min was measured, and the inhibition at each concentration of antibodies was expressed as a percentage of the maximal platelet aggregation in the absence of c3C7 or 3C7. Data were presented as the mean±SD of at least three different determinations.

Jiang, A., Zhang, W., Wu, Q., Jin, W., Tang, Y., Zhang, J., & Liu, J. N. (2014). Construction and characterization of a novel chimeric antibody c3C7 specific for the integrin αIIbβ3 complex. Applied microbiology and biotechnology, 98(1), 105-114.

Figure 7 The cross-reactivity of c3C7 with anti-mouse IgG antibody by ELISA method.

CHO/αIIbβ3 cells were grown to 95 % confluence in the 96-well culture plate. After fixing with glutaraldehyde and blocking, the cells were incubated with 1 μg/mL c3C7 or 1 μg/mL 3C7. After washing, HRP-conjugated goat anti-human IgG Fc, HRP-conjugated goat antimouse IgG Fc, HRP-conjugated goat anti-human IgG H+L, and HRPconjugated goat anti-mouse IgG H+L were used to detect the primary antibody, respectively. TMB substrate was used for color development. The OD of each well was read at 450 nm. The data are expressed as means of duplicates±SD

Jiang, A., Zhang, W., Wu, Q., Jin, W., Tang, Y., Zhang, J., & Liu, J. N. (2014). Construction and characterization of a novel chimeric antibody c3C7 specific for the integrin αIIbβ3 complex. Applied microbiology and biotechnology, 98(1), 105-114.