I am doing ADCC experiments with human sera, using virus-infected epithelial cells as targets and Jurkat Lucia NFAT CD16 cells (Invivogen) as effectors. In the experiment, I seed a 96-well plate with epithelial cells and infect them with the virus, and a control plate of uninfected epithelial cells. After 5 days of infection, I remove the cell supernatant, add serial dilutions of the sera, and incubate the plates for 4 hours before adding the Jurkat cells. The final target:effector ratio I am using is 1:10. After incubating the plates overnight and reading the results in a luminometer, I see that the RLU signal in the uninfected plate is higher than in the infected plate (including the infection-specific positive controls). What could be causing this non-specificity signal?
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