I am trying to calibrate an antibody which on its data sheet specifies it works for parafin IHC. On the datasheet it says it should work with PH-6 HIER method is recommended.
I have used Sodium citrate buffer in PH-6 in the microwave for ten minutes. It did not work. What other methods of unmasking should i try to get a results with this antibody?
To remove the confuse and masking for Antigens , I suggest you to use retrivale solution with heat
This masking result from cross linking between a.as with sorrounding Antigen(epitopes)
For unmasking epitopes of formalin-fixed tissues you can use either pretreatment with citrate buffer (pH6, up to 20min at 100°C), EDTA buffer (pH8, up to 20min at 100°C) or protease treatment (e.g. proteinase K for 30min at 37°C).
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