Usually with IHC protocol, I wash before and after H2O2 with TBS buffer, recently I found some protocols use dH2O instead of the buffer, which one is better?
When you ask what is better, what does this mean in the histochemical sense? It means that your signal to noise is optimal ie you are getting good staining. It would be nice to eliminate unnecessary buffer washes of possible,but sometimes empirical observation must be made.
If you are processing paraffin sections, you will have gone through xylene and then through graded series of ETOH. So you are now in a Low ETOH solution. Some protocols then go to the H202 in a methanol solution or have a H2O rinse before going into a dilution of the H202. However, once you are out of the H2O2, and are going into the blocking sera, you may wish to rinse with buffer.
For antigens which are H2O2 sensitive, an alternative method is used. The H2O2 incubation is done after the biotinalated secondary (if you are using the ABC method). In this case, a good Buffer wash both before the H2O2 and after it are necessary. Good luck
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