Tips for eliminating background when staining mouse tissue with a mouse monoclonal antibody.
Staining of mouse tissue using mouse antibody is a complicated process as high levels of background are often observed. It is notoriously difficult to eliminate this background.
Much of the background is caused by secondary antibody binding to endogenous mouse IgG in the tissue being stained, and to Fc receptors on B cells, plasma cells and macrophages.
Abcam offers a robust one-step kit to allow use of mouse antibody on mouse tissue. ab127055 - Mouse on Mouse Polymer IHC Kit is a polymer based system that provides increased sensitivity, time saving and detection simplicity.
Alternatively, here are a few tips to try and reduce background should you choose not to use Mouse on Mouse Polymer IHC Kit.
Blocking of endogenous IgG
Fc receptors are present on several cell type such as macrophage, and monocytes and can bind to the antibody and give additionla background staining. We recommend using F(ab) monomeric secondary antibodies to help reduce background.
Other tips that can be used to decrease general background: