Before, I was incubating the cells with [3h]Adenine overnight and thereafter separating cAMP (from ATP,ADP and AMP) by sequential chromatography (using Dowex and alumina columns) . This method was efficient and also cheap!
Unfortunately the company (PerkinElmer) that we used to buy [3H]Adenine from them, the only producer of this compound in the world(!!!), is not producing it so often anymore (they produce it only twice a year). For example now (May2017) I have to wait until September to receive [3H]Adenine!
I am looking for an alternative method which the “DIRECT” detection of cAMP assay in that, is independent of any other molecule’s function (e.g. luciferase) and doesn’t require any transfection (e.g. PGL4).
cAMP can be detected by a competitive ELISA kit from cellular extracts.
here is a link (there are a lot of other products):
https://www.caymanchem.com/product/501040/image
Just a short question. What is the problem of the half-year schedule? 3H is a slow decaying nuclid (half-life over 10 years).
Dear Cornelius,
Many thanks for your reply. Hartmann Analytic had it.
Dear Attila,
Thank you for your reply. Actually the problem was that we already run out of it and had to wait for few month! But fortunately Hartmann Analytic still provides it as mentioned already.
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