immunofluorescence, False Positive Nuclear Staining in Hydrogel-Embedded Breast Cells?

29 June 2024 1 301 Report

I have primary breast cells embedded in the hydrogel. When stained with EpCAM without the hydrogel, the staining was correctly localized to the membrane as expected! However, after embedding in hydrogel and using the same protocol for immunofluorescent, we are now seeing nuclear staining. Have you encountered this issue? Any recommendations on how to resolve it?

Howard Vindin

Are you sure that this is an artefact? If the antibody epitope is on the intracellular domain it may be genuine signal.

This paper talks about its translocation:

https://www.nature.com/articles/ncb1824.pdf

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