Double cytokine staining of cells

Hi all,

has anybody intracellularly stained human/primate cells for IFNgamma and
IL-2 as well as a typical surface phenotype stain all at once?

For instance, I am staining T cells with CD4-FITC, CD3-PE, CD8-PerCP and
IFNgamma-APC. I would like to look at co-production of IL-2 but cannot find
the right conjugates. All the antibodies are from BD, and the IL-2 is only
available in FITC and PE. The only other flurochromes I'm able to fiddle
with on our LSR are PE-Cy7 and APC-Cy7, but the surface stain antibodies are
only available in FITC, PE and PerCP.

If anyone has successfullly analysed CD4 and CD8 T cells (preferably human
or primate, not mouse) for IFNgamma and IL-2, I would like to know which
antibodies and flurochromes you've used and where they can be obtained from?

I prefer not to analyse CD3+CD4+ and CD3+CD8+ in separate tubes as I have a
very large sample number with up to 7 different antigens and would then have
to analyse double the number of tubes that would be more than 500 tubes at
any given time. Any advice is appreciated.

Thanks all


Socheata Chea

Department of Microbiology & Immunology
The University of Melbourne
Parkville, VIC, 3010, Australia

Phone: 61 3 8344 0073
Fax: 61 3 8344 3846
Email: schea@unimelb.edu.au