Antibody-dependent cell-mediated cytotoxicity assays (ADCC)Antibody-dependent cell toxicity was determined using the flow-cytometry based method employing the principle of live cell discrimination (Srivastava et al., 2013).Dispense 5.0 x 104 labelled target cells in 50 μl RPMI 1640 media in each well as per the layout given bellow in round-bottomed 96 well plate in duplicate. #####%%%%% output "duplicate" are not used
{"action": "determine", "output": "duplicate", "reagent": ["Antibody-dependent"], "device": ["flow-cytometry"]}
A total of 5,000 target cells acquire. #####%%%%% output "live effector cells (no dye), 7-AAD only" are used in line6
{"action": "acquire", "output": "live effector cells (no dye), 7-AAD only", "reagent": ["5,000 target cells"]}
Determine percentage cell death by software analysis of four identifiable cell populations, live effector cells (no dye), 7-AAD only, live target cells (PKH-67 only) : ﻿0,1,2,3,4,5,6 1,2,3,4,5,6,7 Unstained Target Cells,Unstained Effector Cells,PKH67 Stained Target Cells,7AAD #####%%%%% output "percentage cell death" are not used
{"action": "determine", "output": "percentage cell death", "reagent": ["live effector cells (no dye), 7-AAD only"], "device": ["software analysis"]}