Is Peroxidase kit (Biotinylated secondary antibody followed by ABC reagent incubation) used in IHC light sensitive? Means will it show reduce signal after DAB if we exposed section to light during secondary antibody and ABC incubation?
Yes HRP is light sensitive. It is important to avoid excessive exposure to light, Therefore using dark moisture chember is very useful during these steps.
What sort of photo-reactions/side-reactions one would expect in the presence of light ?
No, the HRP is not light sensitiv. I worked with the ABC kit more then 20 years and I couldn't observe that light reduce the staining intensity of the following DAB reaction. Importen is that you prepare the ABC solution minimum 30 min before you apply them onto your sections. This time is nacessary for the complex building between Avidin and biotinylated peroxydase. If you don't take care about this you will get a result which is less intensive. Evan the DAB complex is stable over years if the sectons are mounted in DPX or other xylenen soluble mounting medium.
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