Use of horseradish peroxidase to block nonspecific enzyme uptake in immunoperoxidase microscopy.
Intrinsic tissue peroxidase activity can be more or less successfully destroyed by methanol-H2O2 treatment. It has been found, however, in our laboratory that horseradish peroxidase (HRP) coupled to antibody will bind to some tissue components on a nonspecific basis and remain to take part in the histochemical stain. This contributes considerably to the background. This difficulty can be largely overcome if the tissues are pretreated with a solution of horseradish peroxidase which binds with nonspecific tissue sites. The adsorbed enzyme, along with the intrinsic peroxidase, can then be successfully inactivated by methanol-H2O2 treatment. By this method of blocking, there is considerable reduction in background staining.
D006651:Histocytochemistry; D006735:Horseradish Peroxidase; D006861:Hydrogen Peroxide; D007124:Immunoenzyme Techniques; D007136:Immunoglobulins / Q000032:analysis*; D008198:Lymph Nodes / Q000276:immunology*; D000432:Methanol; D010544:Peroxidases; D010950:Plasma Cells / Q000276:immunology*
D007136:Immunoglobulins; D006861:Hydrogen Peroxide; D006735:Horseradish Peroxidase; D010544:Peroxidases; D000432:Methanol