Supplementary Figure 9: Assessment of specificity of polyclonal antibody to human GPR15 and comparison of staining to that of an anti-GPR15 monoclonal Ab.
From: Role and species-specific expression of colon T cell homing receptor GPR15 in colitis
Assessment of specificity of polyclonal antibody to human GPR15 and comparison of staining to that of an anti-GPR15 monoclonal Ab. (a) L1.2 cells stably transfected with the human GPR15 ORF and C-terminal GFP tag were incubated with 1 μg (black line) or 0.5 μg antibody (dashed), or were unstained (gray shade; left panel). Polyclonal antibody binds GFP+ cells in human GPR15 transfectant cells (middle panels). L1.2 cells stably expressing human GPR15 (black line), mouse GPR15 (dashed) or control vector (grey shade) were stained with 1 μg antibody, and there is greater binding of antibody to human transfectants (right panel). (b,c) Similar staining of colon CD4+ T cells by polyclonal (b) and monoclonal (c) antibody to human GPR15. Lymphocytes isolated from human colon were activated and incubated with GPR15-specific polyclonal (or rabbit IgG isotype) or monoclonal GPR15 antibody (or mouse IgG2B), Foxp3 and cytokines. Cells are gated on CD4+ CD45RO+ memory cells.
