CD107a staining
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- Put the following together in a round bottom 96 well plate
1x106 splenocytes (100ul)*
1 ul of 1:100 peptide (5ug/ml)
1ul CD107a-FITC antibody
- Spin down at 500xg,1min and incubate 1hr at 37C
- Add 100 ul of 1:750 golgi-stop (monensin) and incubate 4-5 hours at 37C
(Do not use golgi-plug (brefeldin))
- Wash cells 2x w/FCWB and stain with the desired antibody.
* Recommended cell #’s (1:2 E:T ) if using target cells
T Cell Clone: 5x105/96 well
Target cells of peptide loaded splenocytes: 106/96 well