Phosphoflow: I am working on cancer cell lines looking for phosphoproteins by flow cytometry. Any advice?

28 December 2013 1 10K Report

In flow cytometry single histogram of a single phosphoprotein splitted into two peaks. Even that is a stable cancer cell line. Why is this?

Paulo César Martins Urbano

I didn't get what you're looking for... but if I understood correctly you have a histogram where the x-axis is the phosphoprotein (which one?) and you saw two clear separate peaks, therefore you may assume that one peak represents the cells that do not present phosphorylation (the peak to the left) and the peak to the right represent cells expressing the phosphorylation of interest. Use unstimulated and unstained cells to see clearly the difference between positive and negative cells. Please could you add an image and detail a bit more your experimental setup? Stimulation, cell line, phosphoprotein of interest.

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