Staining Intracellular Cytokines
Francesco Borriello
Friday, 28 August 2009 07:28 UTC
Hi all,
I’m going to measure cytokine production by human lymphocytes via flow cytometry and I have some questions for you.
Do I have to block Fc Receptors before fixing and permeabilizing cells? How can I do this?
What is the lowest number of cell I can start with?
Thank you in advance.
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Replies
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Hi Francesco,
I don’t block Fc receptors in my protocol, and people I’ve spoken to have only done this when using mouse cells. I generally do all my setup and staining in a 96-well V-bottom plate, and I aim for 100 000 to 300 000 cells/well. One of my colleagues has gone down to 10 000 cells/well out of desperation, but I wouldn’t use less than 50 000 cells/well in my own experiments!
Good luck!
Michelle
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Hi Michelle,
thank you for your answer, it has been very useful!Best wishes
Francesco
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