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Optimize your experiments
Fig. 1. IHC-frozen mouse intestine cryosections stained with anti-F4/80 (MCA497; red) and anti-PLVAP (MCA2539; green) antibodies. PVLAP stained blood vessel endothelium surrounds F4/80-labeled macrophages in the lamina propria. Nuclei were counterstained with DAPI.
Download the tips and hints for your IHC-paraffin experiments
View the F4/80 minireview
As with every experimental design, the inclusion of positive and negative controls in your immunohistochemistry (IHC) staining protocol is essential. Because IHC is a rather long and complex procedure, thinking of controls early does not only save you time but also eases the troubleshooting process.
The main purpose of the listed control types is to verify that the observed antibody staining is specific.
Crucial Controls and Tips for IHC Experiments - Optimize your experiments
It is also important to block endogenous peroxidases and phosphatases prior to using alkaline phosphatase (AP) / horseradish peroxidase (HRP) antibody conjugates. For blocking endogenous peroxidase activity Bio-Rad offers ready to use peroxide blocking reagent (BUF017B).
When using avidin/biotin or streptavidin/biotin detection systems it is also important to use an avidin/biotin blocking system to minimize non-specific staining caused by endogenous avidin, biotin present in the tissue. For this specific purpose Bio-Rad offers ready to use avidin-biotin blocking system (BUF016).