I am trying to measure cGMP in H9C2 cells. The data I get are out of the calibration curve. Does anyone has an experience or idea how this could be improved? I already increased cell number and it didn't help so much.
May be there is an alternative way of cGMP measurements in cells?
Thank you in advance!
Hii Sofya,
I have used Radioimmunoassay to measure cGMP levels in HEK293T cells. To my experience that is the most sensitive method and you get pretty clean data. However, standardisation of assay may take some time.
However, there is one more option that might be easy for you to follow. You could modulate the standard curve itself. Example, if the data that you are getting is off towards the lower concentration point of your standard curve, then you could try shifting entire standard curve in the same direction. Which means, if the linear region of standard curve falls within micromolar range and the concentration of your sample is in nanomolar, then try reducing the concentration of standard curve such that the linear range falls within nanomolar concentrations.
Hope this could help.
Good Luck.
Hi Insha,
Thank you for your reply!
If I move the standard curve towards the, let's say, picomolar concentration, the sensitivity of the kit won't be reliable due to the kit sensitivity, right? Actually I took one more standard but still the cGMP concentration was out of the curve.
Sofya
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