3 replies to this topic

[irisgzz]

Hi guys!

This might seem a stupid question, but just to be sure!

I want to test a couple of inhibitors in vitro to check expression of genes related with immune response in my cells. I know the IC50 value of these compounds from an assay with 2x10^5 cells/mL, and I have some data about previous titration of this inhibitors (using different concentrations of the inhibitors on 2x10^5 cells/mL to find what is good to inhibit my target enzymes). So, lets say that 1µM is a concentration that is good to inhibit my enzymes without finding cytotoxic effects on my cells.

Now, the thing is that I have to stimulate cells to check for immune response (with LPS, TNF) and see if my inhibitors have some effect. The protocol that I have for stimulation is for 1x10^6 cells/mL. If I would like to add my inhibitors directly to these cells on 1x10^6 cells/mL together with my stimuli,  would it be valid to say:

- 1µM of inhibitor for 2x10^5 cells/mL --> for 1x10^6 cells/mL (5x more cells), then 5µM of inhibitor?

Thanks a lot in advance!

[pito]

1µM for 2x10^5 cells/ml


1µM for 2x10^5 cells/ml
   II
   II  :  2x10^5
   II  X 1x10^6
   V
? µM for  1x10^6 cells/ml




Use simple math: for 1x10^6 cells/ml it is 1µM divided by 2x10^5 and multiplied by 1x10^6 cells/ml, which gives you: 5 µM , or indeed as you allready noticed, its 5 times more cells.. so indeed multiply by 5.

Edited by pito, 16 April 2012 - 01:15 AM.

[irisgzz]

pito, on 16 April 2012 - 01:13 AM, said:

1µM for 2x10^5 cells/ml


1µM for 2x10^5 cells/ml
   II
   II  :  2x10^5
   II  X 1x10^6
   V
? µM for  1x10^6 cells/ml

Thanks a lot! = )


Use simple math: for 1x10^6 cells/ml it is 1µM divided by 2x10^5 and multiplied by 1x10^6 cells/ml, which gives you: 5 µM , or indeed as you allready noticed, its 5 times more cells.. so indeed multiply by 5.

[bob1]

I would say that the amount of compound present in the solution at 1 uM/L is likely to be enough to work for any number of cells, so you shouldn't need to adjust it.