I have two antibodies that are staining well in IHC but not in IF for tissue sections, when using the same concentration and antigen retrieval method for both. I also know the secondary works well.
As far as I know, IF works on frozen sections (antigen retieval not needed on frozen sections), try that
Hi Lucia Marinas del Rey
I agree with Subhash C. Juneja , While IHC typically is performed in formalin-fixed, paraffin-embedded tissue, IF needs to be done with cryotome sectioning (frozen or PFA, sucrose fixed) where an antigen retrieval step is not required.
Hope it helps,
Thanks,
Hello immunohistochemistry IHC can create primary and secondary antibodies,hope you know thermo fisher apats enough wit your role protocol , serially
Fix freshdisseccion overnight
Rinse with running tap water
Dehydrated the tissue 4 times of 100pp alcoholic
So on
But b cells
When immune fluorescence if uses fluorescence signal not for both like faded straining when IHC staining is parmanent and not faded
You have to differentiate between paraffin (ffpe) sections and frozen sections. If both tissue types are formalin fixed and the antibody ist suitable for formalin fixed tissue , then it should work but you have to find out the optimal dilution for your primary antibody, because for IF IHC you need higher concentrations. If the antibody is only used for native (unfixed tissue) and you like to use it on fixed tissue then I would recommand to look for another antibody.
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