How to calculate pg/ml/mg for tissue culture? - (Sep/12/2013 )
I need an advice how to calculate final cytokine concentration in tissue culture in pg/ml/mg. I cultured tissue explants about 200mg per 700ul media. Then I collected supernatants and measured tnf production by ELISA (100ul of supernatant per well). My final result from this assay is in pg/ml but I need to calculate them with relation to mg of tissue.. How can I do that?
I tried to express my tissue culture concentration as 280mg/ml - but what I should do after that? Should I devide my ELISA result or multiply? A little bit confused..
How I would do it is to calculate how much (total) cytokine was produced - i.e. take your concentration and multiply by the volume (keep units consitent, e.g. if you have pg/ml then the volume needs to be in ml...)
Then you have a total cytokine and you can work out how much was produced per mg tissue. Units will not be pg/ml/mg, but pg/mg!
thank you very much for your answer!
you mean that:
if my elisa result for TNF-a is 200pg/ml, I should multiply this ammount = 200pg/ml x 0.7ml (culture well volume) = 140pg. And then I should devide it to 200mg ot tissue (tissue mass in a well) = 140pg / 200mg = 0.7pg/mg
am I right in my calculations? this way definitely correct to know how much was produced by mg..
I was just wondered about these units, because in papers with the measurment cytokines in tissues they put in their graphs "pg/ml/mg".. but if the way above is correct I could use it!
thank you again!
I don't know how the papers do it - I don't work with cytokines, but in terms of science units like pg/ml/mg make no sense - think about it picograms per ml (of what) divided by mg of tissue? The tissue mass and ml are unrelated and the volume is irrelevant other than as a normalizer to get the total cytokine production.
The way I suggested above is how I would do it, but if you followed your papers you would get 200 pg/ml/200 mg = 1 pg/ml/mg.
thank you for your answer!! I think you are right.. I will calculate by your way! it is more reasonable..