Question

we have two drugs used ( Nimesulide and Celecoxib) to inhibit the expression of cox-2 in three cell lines at different concentrations(1,10,20,30,50,70,100) microliter. How to preform the apaptotic essay?

Answer 1

Ans: Protocol used for performing apoptotic assay is explained below-

1. Cells were grown till the logarithmic phase

2. Treat cells with Nimesulide and Celecoxib at different concentrations (1,10,20,30,50,70,100) and incubate for 24 h

3. Harvest cells and centrifuge it

4. Collect cells, wash it using PBS, and resuspend it in 500 µL of 1X binding buffer.

5. Add 5 µL of annexin V-FITC and 5 µL of propidium iodide

6. Incubate cells in dark at room temperature for 5 mins

7. Quantify it using flow cytometry or fluorescence microscopy.