I have been doing some flow cytometry on T cells and i am trying to find the
balance of T helper cell one and two subgroups by measuring the amount of il-4
and IFN GAMMA, i have been having trouble getting any IL-4 staining with a FITC
anti human IL-4, on CD4 positive cells, so to test my antibody i did the test
with a different antibody to IL-4 conjugated with PE, but as i only had a FITC
CD3 antibody t wasn't specific for T helper cells. I found i had good staining
for IL-4 both on CD3 positive and negative cells, and i am having trouble
finding out what group of cells are CD3 negative and produce IL-4 on
stimulation. I have gated on lymphocytes only on the flow cytometer so they are
not anything else such as mast cells.
i would be grateful for any suggestions, and would like to know if any-one else
has had trouble getting IL-4 production from activated T cells from healthy
volunteers.
Anna
