I am interested in studying priming-associated gene expression in naive human CD8 T cells. I would like to find the best conditions for their short-term in vitro activation, so that their priming process is as close to physiological one as possible. I am planning to use plate-bound anti-CD3 and anti-CD28 antibodies. What is the concentration of anti-CD3 and anti-CD28 antibodies that you would recommend? Would you prefer soluble anti-CD28 to plate-bound one? Would you use IL-2?
Thank you!
Hey Maarja,
In case you're interested in a different approach, I can recommend the CD3/CD28 Streptamer® Kit (https://www.iba-lifesciences.com/cd3-cd28-streptamer-kit-human/6-8900-050). Instead of high affinity antibodies, this approach uses low-affinity Fab fragments that are first multimerized on a backbone and then just added to the cell culture well. The advantage here ist that the multimerization is disrupted after biotin addition and the stimulation of your cells will stop immediately. This might be of interest to you, because you only need short term stimulation. The Fab fragments will completely dissociate from the cell and all reagents can be removed by washing. So you don't have any antibodies stuck to your cells that might affect your analyses.
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