Optimization of Mouse-on-Mouse Immunohistochemistry by Utilizing Fluorescent-dye Conjugated Secondary Anti-Mouse Antibody

Abstract

The application of mouse monoclonal antibody for immunostaining the mouse tissues results in a high rate of background noise because of the interaction of the secondary antibody with endogenous immunoglobulins and other immune components. The most advised blocking strategy for the mouse-on-mouse immunostaining is the use of anti-mouse Fab fragments. Nevertheless, the commercial kits containing Fab fragment are costly and unavailable in many research laboratories. In this study, we provide evidence showing the potential of the fluorescent-dye conjugated secondary anti-mouse antibody for reducing the background noise in the mouse-on-mouse immunohistochemistry. Furthermore, our findings demonstrate the inadequacy of goat serum/protein-blocking solution alone as an immunohistochemistry blocking system for reducing the background noise.