[HISTONET] Antigen retrieval paraffin skin samples

[Nicholas David Evans]

Dear all,

 

Might anyone be able to offer their advice on the best method to retrieve
antigens on paraffin embedded skin sections? I am immunostaining for
epithelial cytokeratins, and I have block-to-block variation in the
quality of the staining - some block give consistently good staining while
others giving negligible staining. The tissues were prepared by fixation
in 4% PFA for 24 - 48 hours followed by dehydration and embedding. I
currently use 15 mins Ficin digestion. I suspect that the differences may
be accounted for by variability in the length of time samples were fixed
in PFA and then 70% ethanol.

 

Best wishes

Nick