>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 Visia, We have had good success using the Caltag antibody, Cat MHBCL01-4, which is FITC conjugated. We use it for flow cytometry phenotyping of our human-BCL-2 transgenic mice by intracellular staining and it works great. They also have a biotin conjugate of the same clone available, although we have not tried it. Check with Caltag to see if it has been tested for Westerns. Randy T. Fischer NIA/NIH 5600 Nathan Shock Dr. Baltimore, MD 21224 (410) 558-8452 (410) 558-8237 (fax) fischerr@vms.grc.nia.nih.gov
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