Re: PE-Cy5 or PER-CP
(no name) ((no email))
3 Dec 96 10:48:13 EDT
We use primarily PerCP (exclusively a BD product) for lymphoid subset
evaluation in marrow, peripheral blood and cord blood. The fluorescent
intensity is between 100-1000 fluorescent units (depending upon mAb), 0%
spectral compensation between FL2 and FL3, and good discrimination between
positive and negative. While the CD34PerCP and HLA-DR(II)PerCP mAb have an
intensity mean of 120 fluorescent units, as with the FITC conjugates, it is not
strong enough for the earlier progenitors. For our stem cell subset
evaluations we are using CyChrome (PharMingen Inc) a PE-Cy5 tandem in
combination with CD34PE which gives us a bright signal, with some spectral
cross-over (1-2% in FL2-FL3), but not as much as we noticed with the TriColor
(Caltag) tandems (4-5% in FL2-FL3). Another source of PE-Cy5 antibodies is
Immunotech, which gave us similar results to the PharMingen tandems. When
setting up our three color panels, we evaluated various fluorochrome
combinations to determine which combo gave us the best discrimination for the
subset were were evaluating. Good Luck!
April Durett
Technology Transfer - Flow Cytometry
Blood & Marrow Transplantation
Univ Texas MD Anderson Cancer Center
adurett@notes.mdacc.tmc.edu
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