Product: Brilliant Violet 421™ anti-human CD25

Catalog No.: 302630

Anne Rissiek, Dr. Universitätsklinikum Hamburg-Eppendorf (UKE)

CD25 Brilliant Violet 421™ to Discriminate Human Tregs in Whole Blood
Overall:	Product Quality:	Ease of Use:
Using this antibody it is possible to discriminate human Tregs nicely.

Experimental Design

Application:	Flow Cytometry
Cells used:	Human EDTA blood
Brief Protocol:	1. 100 µl EDTA blood plus 50 µl antibody cocktail (in FACS Buffer), incubate for 30 min at RT. 2. Lyse red good cells by adding 1 ml Lysis Buffer, incubate for 10 min at RT, wash 2x with 1 ml PBS, 450 g, 5 min, trash supernatant. 3. Resuspend cells in 200-300 µl FACS Buffer, FACS analysis. The CD25 antibody was titrated and applied at a final dilution of 1:500.
Results Summary:	In combination with anti-human CD127 Brilliant Violet 421™ (clone A019D5, BioLegend) this antibody allows an excellent discrimination of human Tregs.
Additional Notes:	Gating strategy: Lymphocytes --> Single cells --> T cells (CD3+) --> not gd T cells --> CD4+ --> Tregs (CD127loCD25hi)

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