Hi

one day sice 20 years (2000), I was studying key apoptosis and cell cycle regulatory proteins in human colorectal cancer ( CRC ) biopsies as well as 5 CRC cell lines.

I am the first one in this world to realize that Bcl2 expression is nuclear during apoptosis and mitosis in cancer cells by immunohistochemestry ( Elkablawy, 2001 Journal of Pathology).

I had to confirm this expression and localization by other methods or techniques.

I used Immunoflurecence staining with Confocal Laser Staining Microscopy (CLSM).

Then I used analyze PC and Irfan V programms to build up a 3D images and vedios with doing transparancy to see the localisation inside the nucleous from different planes.

Finally I used a techique to separate the nucleus from the cytoplasm and did Western blotting for both of them separatly.

Also I did flowcytometry to study the expression in the different stages of the cell cycle.

Good Luck

MOHAMED ELKABLAWY. Preferred source ... Bcl-2 and apoptosis in colorectal cancer : a molecular and quantitiative analysis. 2002-01 | dissertation-thesis.

Queens University of Belfast, N. Ireland, Uk.