BD Biosciences
TCR gd BUV563
748993
GL3
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I used this antibody to identify gd T cells in a panel also analyzing conventional T cells, B cells, monocytes, DCs and NK cells in C57BL/6 mice in different organs.
Flow Cytometry
C57BL/6 splenocytes
1:100, 20 min, 4°C, in 50ul, in the dark
Fc Block
Fix/Perm buffer, 20 min, 4°C, in 70ul, in the dark,
20 min, 4°C, in the dark, in 50µl Perm/wash
FLow Cytometry
The antibody worked well to identify gd T cells in multiple organs of C57BL/6 mice. Similar to all other BUVs, the antibody is a bit pricey; I use it at 1:100 in 50ul, which makes it reasonable to use. The fluorochrome BUV563 is not super bright, but brightness is sufficient to identify these cells and spillover into neighbouring channels is minimal. I used the marker in combination with a-CD3 (clone 17A2) FITC, which worked well to identify this T cell subset. I would recommend this antibody/ fluorochrome combination. In the following, you will find my staining protocol in short. Incubation steps were performed at 4°C in the dark. 1.Count and adjust splenocytes to 1.5E6 cells, plate in a 96-well plate. 2.If required, stain cells with viability dye. Wash. 3. Stain with extracellular master mix for 20 min. Wash 2x. 4. Fixate/ Permeabilize with buffer of choice, depending on application. I used ThermoFisher FoxP3 fixation/permeabilization kit for 20 min. Wash with perm/ Wash 2x. 5. Perform intracellular staining for 20 min. Wash 2x. 6. Resuspend in FACS buffer and acquire as soon as possible. Here, data was acquired on a 5L Cytek Aurora.
N/A
Clear signal, good correlation with CD3.
Works well, easy to use. A bit expensive, but at 1:100, you can stain a large number of samples.