I have tried to activate my jurkat cells using the dynabeads human t-activator cd3/cd28 beads. Was wondering how long do you have to incubate your cells with the beads before preforming a flow cytometry ?
Best/ M
I activate Jurkat cells with soluble anti-CD3. I look at activation markers after overnight stimulation or 24h. You can look as early as couple of hours, but then you will look at the surface expression of already synthesized protein rather than neosynthesis. Though, in my experience early and late time points gave the same trends
Hi Mahdi.
For short time activation, the cells can be lysed while they are still on the beads for molecular assays. Between days 1-3 the Dynabeads form strong clusters with the cells, thus it is hard to remove the beads in this period without losing a lot of the cells. After 3-4 days in culture, most of the Dynabeads have detached from the cells and you can remove the beads on a magnet prior to flow. For more questions, contact me at [email protected]
Hi, I would like to ask if we are using Dynabeads CD3/CD28 to activate Jurkat cell, what cytokines can be released?
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