PMA/Iono - (Sep/19/2009 )

Hi,
I'm going to stimulate human CD4 T lymphocyte with PMA/Iono for intracellular cytokine staining
and I have some questions:

1) What concentrations of PMA and Iono do I have to use?
2) Is it enough to stimulate cells for 4 hours?
3) Do I have to add Bleferdin A at the beginning or in the middle of stimulation?

Thank you!!!

Dear Jay,
Thank you for your answer!
I have another question for you...Do you know any other way to stimulate CD4 T lymphocytes for intracellular cytokine staining besides PMA/Iono?