I am staining for E-cadherin (membrane) and vimentin (intracellular) marker for my EMT study. Anybody has a working protocol where I can do these simultaneous staining on 1 sample.
Or is it better to have two coverslips of the same sample and stain them separately?
Dear Zubaidah,
You can show both membrane and intracellular markers at same time by double staining with IHC. Your targets have no problem for performing. Vimentin always located at intracellular compartment. E-cadherin is a membrane member, but it is translocated from cytoplasma.So, you can find a little bit amount in cytosol. But, there is no problem. You have different choice for staining these markers:
1. After fixation, you can add primary antibodies for both markers and incubate for a sufficient time. Wash. Adding secondary antibodies and realize other steps according to manufacturer advice. (simple method for these markers).
2. Two steps staining for primary antibodies.
3. Choose different species primary antibodies and also secondary antibodies.
.....
Good luck.
Have you tried to stain them seperately, but with the same pretreatment to test, if they work?
I do routinely E-Cadherin and vimentin on FFPET with HIER on both antibodies.
Questions: do you stain Immunofluorescence or chromogene IHC? cells? tissueslides? fixed? cryosections? .....
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