protocol that we used: 1. Soaked the immunofluorscence slides in PBS
2.Rehydrate further with ethanol series : 2 times of 5 mins with 100%, 95% , 70% and 50% of ethanol volumes for 5 mins separately And finally 5 mins in double distilled water.
3. Incubate slides for 1 hour and 2 hours in 40 ml SDS-Glycine pH 2.0 buffer at 50C with gentle shaking at160rpm in bug shaker,
I used the following protocol for destaining immunohistochemistry:
1. Decoverslip slides in distilled H2O (in TBST 5 min)
2. Destaining-dehydrate-rehydrate:
2 min in DiH2O
2 min - 70% EthOH
10 min - 95% EthOH
2 min - 70% EthOH
2 min - DiH2O
3. Antibody elution - incubate sections in preheated at 50C Glycine SDS buffer pH2.0, agitating for 1-3 hours (depends on persistence of antibody)
Wash in DiH2O 10 min
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