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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