I want to stimulate jurkat cells with PMA and ionomycin. Whether it is necessary to add calcium to the medium. if needed, How much concentration should I use?
Hi Wansheng,
If the idea is to provoke a long-lasting intracellular calcium signal that will evoke a cellular response, then yes you need to have extracellular calcium in the medium. Most people use 1.5 - 1.8 mM extracellular calcium.
Without extracellular calcium, ionomycin will evoke on a brief calcium signal (due to release of finite intracellular calcium stores; mostly calcium in the endoplasmic reticulum).
Best of success with your experiments.
Martin
Do you mean your cell culture medium (RPMI for example)? If yes it should already have around 0.4 mM CaCl2; you can check its formulation.
Dear Martin
thank you so much and your advice is very helpful. I will try the method you told me.
wangsheng
Dear J. Ashot Kozak
thanks a lot. I have checked my cell culture medium and it has 0.4mM Ca(NO3)2 .I will consider this impact on my experiment.
Best wishes for you
wangsheng
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