[Cytometry] PE binding
davidspurdueemail at gmail.com
Sun Jan 22 10:29:15 EST 2012
I recall PE-conjgates such as PE-Cy5/5.5 and maybe PE-Cy7 having the proclivity to non specifically attach to Fc receptors. I wouldn't rule this out until you attempt blocking FcRs.
Core Director MHRI-Houston
Sent from my iPad
On Jan 20, 2012, at 11:19 AM, "Bui, Hanhvy T." <hbui at jhsph.edu> wrote:
> Hello All,
> I'd like your thoughts on a phenomenon I've seen when staining a particular patient's peripheral blood. This HIV- and HCV-positive patient's cells have shown a high degree of phycoerythrin binding, with some of the cells reacting positively with all PE-conjugated antibodies (CD8, CD38, and IgG1 in our current staining panel) to such a degree that the fluorescence is at times off-scale. Since the cells are prepared from fresh whole blood and the hyperfluorescence appears slightly dimmer when the blood is washed prior to staining, I suspect that there may be some factor in this patient's plasma contributing to the unusual PE-binding. The patient has been drawn twice (earlier in the month, and then a year prior), and this staining was seen both times.
> Has anyone observed this phenomenon before or have any explanation to offer? I have some thoughts, but I'd certainly value the flow community's input. I've found in the literature so far only one similar incidence seen on B cells. In my case, however, it appears that all of the cells are binding PE. If you'd like to take a look at the PDF or FCS files, let me know.
> Thanks for your help!
> Hanhvy Bui
> Johns Hopkins University
> Bloomberg School of Public Health
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