Apc conjugated mouse anti human igg fc antibody hp6017

For rat anti-mouse PVRIG monoclonal antibody (Clone 1) binding assay, 2 × 10⁵ 293T-mouse PVRIG cells were incubated with different concentrations of antibodies (Clone 1), and then the binding frequency was detected using APC-conjugated goat anti-rat IgG antibody (Poly4054, Biolegend, San Diego, USA). For assessing PVRIG-PVRL2 antagonistic activity, 2 × 10⁵ 293T-mouse PVRIG cells were incubated with different concentrations of antibodies (Clone 1) and 10 μg/mL mouse PVRL2-hFc fusion protein. APC-conjugated mouse anti-human IgG Fc antibody (HP6017, Biolegend, San Diego, USA) was used to detect the binding frequency of mCD112-hFc fusion protein.
For mouse anti-human PVRIG monoclonal antibody (Clone 2) binding assay, 2 × 10⁵ 293T-human PVRIG cells were incubated with different concentrations of antibodies (Clone 2), and then the binding frequency was detected using APC-conjugated goat anti-mouse IgG antibody (Poly4053, Biolegend, San Diego, USA). For assessing PVRIG-PVRL2 antagonistic activity, 2 × 10⁵ 293T-human PVRL2 cells were incubated with different concentrations of antibodies (Clone 2) and 10 μg/mL human PVRIG-Fc fusion protein. APC-conjugated mouse anti-human IgG Fc antibody (HP6017, Biolegend, San Diego, USA) was used to detect the binding frequency of human PVRIG-Fc fusion protein.

Tumor cell lines were incubated with 40 μg/mL Vγ2 X PD-L1 (Target) or Vγ2 X Null (Null) for 1 hour at 4°C, then stained with APC-conjugated mouse-anti-human IgG Fc antibody (HP6017, Biolegend, San Diego, USA) for 30 minutes at room temperature. The APC positive populations and MFI of the APC channel were determined by flow cytometry. The expression scores were defined by [log₁₀ (Target _{APC positive populations} - Null _{APC positive populations}) + log₁₀ (Target _{APC MFI/}Null _{APC MFI})]/2.