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Measurement of adenylyl cyclase by separating cyclic AMP on silica gel thin-layer chromatography
Haruhiro Higashida1, Kazi Z Hossain, Hiroko Takahagi
1Department of Biophysical Genetics, Kanazawa University Graduate School of Medicine, Japan. haruhiro@med.kanazawa-u.ac.jp
Analytical Biochemistry
|September 18, 2002
Abstract:
A procedure for isolation of cyclic AMP (cAMP) by thin-layer chromatography on silica gel is described. One-dimensional ascending chromatograms were developed using [H(2)O/C(2)H(5)OH/NH(4)HCO(3) (30%:70%:0.2M)] as the mobile phase. This procedure separated [32P]cAMP from other radioactive metabolites of [32P]ATP in up to 19 samples on one sheet (20 x 10 cm) over 40-60 min at room temperature (21 degrees C). This simple and rapid isolation method provides a novel and convenient technique for the assay of adenylyl cyclase.