Staining of Intracellular Cytokines
Posted 27 August 2009 - 04:34 PM
I'm going to measure cytokine production by human lymphocyte stimulated with PMA/Iono in the presence of Bleferdin A 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.
Posted 28 August 2009 - 04:54 PM
If you can start with a million cells, you should be in good shape. You can use 500,000 cells and still get ok results.
Posted 28 August 2009 - 04:55 PM
Posted 28 August 2009 - 11:17 PM
I will start from CD4+CD161+ T lymphocytes, so I won't need to gate on CD4+CD8+.
I was thinking about performing these experiments in 96-well plates, but as far as I know the maximum number of cells that can be loaded into such wells is 2*10^5 - 10^5.
So this number would be too low, right?
Posted 04 September 2009 - 05:40 PM
Posted 08 September 2009 - 04:09 PM
Could you explain me in greater details how to block Fc Receptor on humsn lymphlcytes
Posted 08 September 2009 - 06:53 PM
For mouse cells, BD has a good product. For human cells, there is a few out there.