I am currently using Perkin and Elmer Alpha screen cAMP assay and I have been having issues analysing my data.
I carried out my first experiment using HEK-293 cells and stimulating them with forskolin at concentrations 1x10-4 to 1x10-10 to produce a concentration response curve.
Anyway, from my understanding of the cAMP assay kit the LOWER the signalling produce...the greater stimulation of cAMP and because of this I am having problems plotting my data on a concentration response curve.
In the assay the cAMP you want to measure competes with the biotynylated-cAMP that is a part of reagent fluorescent conjugate. Geenerally more of Your cAMP then less signal you obtain.
https://www.perkinelmer.com/pl/product/alphascreen-camp-detection-kit-1000pts-6760635d
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