Hello Ayyoub Kihel

Stimulating agents such as phorbol-12-myristate 13-acetate (PMA) and ionomycin are useful to activate transcription factors for intracellular signaling and production of cytokines of many different immune cell types.

I would suggest you stimulate murine splenocytes with PMA (50 ng/ml)/ionomycin (1µg/ml) for 5-6 hours at 37 degree C.

Best.

@malcolm nobre thank you

Good day, Ayyoub Kihel

Yes, LPS can be used. The recommended concentration is 10 ug/ml. As for the incubation time, we have done 18 h, but anything from 12-24 h could be used.

Good luck with your experiments.