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CD107a staining


 
  • 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