High background signal – low reproducibility
Antibody selection is one of the most critical steps in getting high-quality flow cytometry results. High background signal from antibodies often compromises it. Antibodies not optimized for flow cytometry can detect signals that are not specific to the antigen resulting in high background signals. One of the reasons for high background signals is non-specific binding of antibodies to Fcγ receptors (FcRs) found on macrophages, monocytes, dendritic cells and B cells. To avoid this type of non-specific binding, FcR blocking reagents are used during antibody staining. However, the Fcγ receptor blocking procedure end up being time-consuming and costly.