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Optimization of Mouse-On-Mouse Immunohistochemistry by Utilizing Fluorescent-Dye Conjugated Secondary Anti-Mouse Antibody Publisher Pubmed



Moein S1 ; Adibi R1 ; Amouheidari A3 ; Gheisari Y1, 2
Authors
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Authors Affiliations
  1. 1. Department of Genetics and Molecular Biology, Isfahan University of Medical Sciences, Isfahan, 8174673461, Iran
  2. 2. Regenerative Medicine Research Center, Isfahan University of Medical Sciences, Isfahan, 8174673461, Iran
  3. 3. Department of Research and Education, Isfahan Milad Hospital, Isfahan, Iran

Source: Applied Immunohistochemistry and Molecular Morphology Published:2021


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. © 2021 Lippincott Williams and Wilkins. All rights reserved.