Many labs are measuring degranulation of T cells by cell-surface expression of CD107 (see Betts et al., J Immunol Methods 281: 65 (2003), Link to PubMed abstract), and it's often desirable to simultaneously detect intracellular cytokines, such as IFNg. Unfortunately, optimization of both of these readouts in the same stimulation is not trivial. I suggest the following tips:
1. Stimulate cells in the presence of 5 ug/mL EACH of brefeldin A and monensin. Monensin helps maximize the CD107 readout, whereas brefeldin A helps maximize the IFNg readout. To acheive these final concentrations without addition of an excessive amount of solvent, dissolve monensin (Sigma #5273) at 5 mg/mL in methanol, then dilute 1:10 in PBS on the day of use, followed by 1:100 dilution into the culture medium. Ditto with brefeldin A, using DMSO as the initial solvent (or use the FastImmune brefeldin A from BD [Link to product description], which comes as a 5 mg/mL stock in DMSO).
2. Use antibody to CD107a (use of CD107b antibody adds only slightly to the signal) in PE [Link to product description] or PE-Cy5 [Link to product description], as these give the brightest signals.
3. Add the antibody, at 10 uL per 200 uL of cell stimulation culture, at the beginning of the activation period, and activate for 5-6 hours. Continue with processing for surface and intracellular staining as usual.