I have used 40,000 cells/well for my cell line cytotoxicity assay in 96 well plates. the volume was 60ul. i have used 10ul of 100uM concentration of drug.
Now I want to do immunoblotting of the same drug in a 6 well plate with a 2mL volume. How do I do this?
Hi,
If I were setting this experiment with adherent cells I would use the cell density/cm2 to give me how many cells to plate out in a 6 well plate. For example, 40,000 cells/well in a 96-well plate will give 125,000 cells/cm2 (40,000 cells divided by the surface area of a 96-well plate ~0.32cm2), then I would multiply this by the surface area of a 6-well plate which is ~9.5cm2 to give me 1,150,000 cells/well in a 6 well plate.
To work out the drug concentration, just work out the volume to add to a 2mL volume as opposed to a 60uL volume.
Hi,
If I were setting this experiment with adherent cells I would use the cell density/cm2 to give me how many cells to plate out in a 6 well plate. For example, 40,000 cells/well in a 96-well plate will give 125,000 cells/cm2 (40,000 cells divided by the surface area of a 96-well plate ~0.32cm2), then I would multiply this by the surface area of a 6-well plate which is ~9.5cm2 to give me 1,150,000 cells/well in a 6 well plate.
To work out the drug concentration, just work out the volume to add to a 2mL volume as opposed to a 60uL volume.
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