I am looking for a-human bcl-2 antibody for Flow Cytometry conjugated preferably to FITC which will be able to differentiate between cells expressing low levels of bcl-2. Antibodies I have checked so far give me a very nice negative background with an isotypic control but they appear to produce some background staining which covers low expression of bcl-2. Cells like HL-60 expressing bcl2 low are stained with almost the same intensity as other cells known from Western blot to give much stronger signal. I have checked few fixation/ permeabilization methods and have got similar results. Have anybody out there got a good correlation between Western and Flow using the same bcl-2 antibody (like DAKO clone 124) or is there a pair of different antibodies which are able to correlate well for both applications (including lower values)? Will appreciate some advice/discussion on the subject. Visia Dragowski BC Cancer Research Center
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