I would like to stain lymphocytes using an anti-IL2 antibody (for example), sort the positive cells using a flow cytometer and isolate DNA for PCR. Is this possible? Do you have some reference regarding this?
Hi,
For IFN-gamma producing cells, a kit from MACS using magnetic beads can isolate viable IFN-gamma secreting cells (http://www.miltenyibiotec.com/en/clinical-applications/clinimacs-system/clinimacs-reagents/clinimacs-cytokine-capture-system-ifn-gamma--product-line.aspx). Probably, there is a similar kit to isolate IL-2 secreting cells.
I remember to read somewhere that is also possible to sort viable cytokine secreting cells using bivalent antibodies (against a surface marker and the desired cytokine) but I do not know any manufacturer for antibodies like these.
I hope that it can help you.
Good luck!
Hi, If you want to sort live cells that secrete the cytokine, try cytokine secretion assays (catch assays). Miltenyi has a series of kits that help detect cells producing different cytokines. Check out the manual for IL-2 Secretion Assay – Detection Kit (PE), human.
For PCR you should isolate mRNA not DNA since there won't be any difference between sorted and non sorted cells in terms of DNA content.
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