Good Antibody to Detect TIGIT on T Cells

Results Summary

With this antibody, I was able to detect TIGIT expression on CD8 T cells during murine experimental malaria. The signal is not super strong, not forming a distinct population of TIGIT+ cells, but rather a “smear”. Accordingly, a bright fluorochrome like PE-Dazzel594 and a high concentration (1:50 in 50 ul) are beneficial. That being said, TIGIT detection is difficult and I can see a clear difference between infected and naïve animals. Additionally, the signal correlates with other co-inhibitory molecules like PD-1, LAG-3 or TIM-3. Accordingly, I think this is one of the best choice to detect this molecule. However, sufficient controls should be used to assure correct interpretation of the staining.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 5 L BD LSRFortessa with the appropriate filters.