In vivo preparation of [32P]cAMP and thin-layer chromatography of cAMP-related compounds
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A simple and rapid method for preparing [32P]adenosine 3′,5′-cyclic monophosphate (cAMP) is described. A culture of an Escherichia coli mutant which excretes cAMP about 150 times faster than does a wild-type strain was incubated overnight with [82P]orthophosphate of high specific activity (e.g., 4000 Ci/mol (1 Ci = 37 GBq)). The [32P]cAMP which accumulated extracellularly was then purified to 99.9% radiochemical purity in less than 4 h by adsorption to charcoal and alumina column chromatography. A two-dimensional chromatography system using a PEI-cellulose plate is also described which should prove useful for studying cAMP metabolism with 32P- or 3H-labeled cAMP or ATP.
2018 ◽
1982 ◽
Vol 47
(03)
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pp. 244-248
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2021 ◽
1993 ◽
Vol 75
(2)
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pp. 559-565
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Keyword(s):
1985 ◽
Vol 249
(1)
◽
pp. E77-E88
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