Antigen retrieval IHC Fluorescence

[Sarah_89]

Hi at all

I am new here so first some words to myself; I am a Bioanalytics student, at the moment doing my bachelors thesis in cancer research focused on molecularbiology.

That brings me directly to my question:

This week I prepared a cell staining (Kuramochi & Ovcar8), with different combinations of AB (KI67, CK7, cl.-cas3 and MSLN). I checked them at the confocal microscope and everything went well, except Mesothelin. There i just have noise & brightfield. I have to check now for a possible antigen retrieval, I know there are different methods with microwave, steamer, autoclave, proteolytics, waterbath or combinations. Is there something I should prefer? (The cells are fixed with 4% formaldehyde, covered with Prolong Gold mounting media and a fixed glass which I can remove if needed). If I use one of the methods, is there any damage for the other antibodies?

 

Thanks for your help!

 

best, Sarah