BioLegend
PE/Dazzle™ 594 anti-human CD366 (Tim-3) Antibody
345033
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Our group studies T cells in the blood of untreated chronic HIV+ individuals. We are constantly expanding and optimizing our T cell panels to measure various T cell subsets, activation, and inhibitory receptor phenotype. We picked Tim-3-PEDazzle because fit it well into a 13-color panel we designed to study T cell exhaustion.
Flow Cytometry
Human PBMC
0.5uL Tim-3-PEDazzle in 200uL PBS with 10^6 cells; 15 minutes; room temperature
None.
BD LSRII Flow Cytometer
We first titrated Tim-3-PEDazzle alone to determine the volume that yielded the best stain index. 1uL yielded the best stain index, however, when we used it in our 13-color panel there was significant overlap between the PE and PEDazzle channels. Thus, we optimized both markers within the 13-color panel and 0.5uL Tim-3-PEDazzle worked the best without reducing the frequency of Tim-3+ cells detected.
0.5uL works well; plays well with other colors
Bleedover into PE, but titrating down and FMOs can resolve this issue.
Tim-3-PEDazzle is a great option for detecting Tim-3+ T cells in a multi-color panel.