Mouse IgG2b (genomx cat# 648401) raised against human TRACp protein as antigen, is also cross reactive to mouse and rat species.
Can this antibody be used to label TRACP in mouse cell line?
Thereafter can fluorescent conjugated secondary antibody be used against primary mouse IgG?
Will the usage of secondary antibody against primary mouse IgG antibody in mouse cell line show non-specific binding?
Cross-species reactivity should not be a problem, as long as the antibody does not cross-react with other proteins (related to TRACP). You are using a Biolegend antibody raised against the entire protein. You will have to make sure there is no cross-reactivity for example by comparing TRACP positive tissues with TRACP negative tissues. For testing secondary, always compare with and without primary in parallel.
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As I understand, the cross reactivity you describe is a mouse-on-mouse (MOM) problem. Although they do not declare what is in their sera, Vector have developed an efficient strategy to circumvent this background staining, by using their MOM kit. We are applying this routinely in our lab. Also, you want to make sure you apply highly pre-adsorbed secondary antibodies to reduce spurious tissue staining, e.g. from Jackson.
Yes, the antibody you are using is a mouse mab. The mentioned MOM kit might work. If not, this mab is delivered purified so you might want to conjugate it to avoid the need of anti-mouse secondary.
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