I'm examining mouse intestines by E-cadherin IHC to determine the effects of an enterotoxin on intestinal integrity. I was surprised to find that the outer layer of rounded cells that have detached from the tissue and floating in the lumen are still stained with E-cadherin antibody.
The cells you stained could be dead and therefore give you false-positive results, so that you should exclude.
Than, attachment to the basal membrane is not dependent on E-cadherin which is a cell-cell adherens protein, but on integrins, of which integrin ß1 is the first I would try.
As you are staining these tissues, it would be worthwhile to also include some tight junction markers such as ZO-1 and Occludin
Wei Ping,
Think about where cadherins are located (adherens junctions). How to cells attach apical/basally? Not cadherins!!
What other cation-dependent adhesion are present in epithelia? Look it up!! (hint: what are desmosomes, hemidesmosomes, tight junctions, gap junctions, etc.)
DIL
Try to find E-cad of rounded cells by western blot using those antibodies. Or your antibody is not specific to the target maybe, so you also can check it by Western blot.
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