Variable expression of antigen in various subcellular locations - Any ideas why? - ICC Staining on Glioma Cells (Jul/30/2009 )

Hi guys,

I have done quite a lot of ICC staining for various antigens on human glioma cells, however one antigen NG2 (cell surface) I have variable amounts of staining for in various subcellular locations - does anyone know of a reason to explain this? I have seen it perinuclear, cytoplasmic and at the edges of the cytoplasm!

Any ideas would be great! Thanks.

Sam

some proteins, such as the estrogen receptor are internalized in response to a signal, which is usually the binding of their ligand (in this case, estrogen).

Gonna go out on a limb here and ask if you see different pattern of staining for other
cell surface proteins?
The reason I ask is that if your cells are intact, they are covered by a membrane. This protein covers the membrane so it should look like its present everywhere.
Picture a basketball flattened and you are staining for something on the surface of the basketball. It will look like it is nuclear and cytoplasmic, but it is merely on the surface of the cell.
Otherwise, maybe the antibody is crappy.