Measurement of adenylyl cyclase by separating cyclic AMP on silica gel thin-layer chromatography.
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.
Higashida
Haruhiro
H
Department of Biophysical Genetics, Kanazawa University Graduate School of Medicine, Japan. haruhiro@med.kanazawa-u.ac.jp
Hossain
Kazi Z
KZ
Takahagi
Hiroko
H
Noda
Mami
M
D000262:Adenylyl Cyclases / Q000378:metabolism*; D000818:Animals; D001345:Autoradiography; D002772:Cholera Toxin / Q000494:pharmacology; D002855:Chromatography, Thin Layer / Q000379:methods; D000242:Cyclic AMP / Q000032:analysis / Q000096:biosynthesis*; D005910:Glioma / Q000378:metabolism; D007545:Isoproterenol / Q000494:pharmacology; D007553:Isotope Labeling; D037342:Pertussis Toxin / Q000494:pharmacology; D010761:Phosphorus Radioisotopes; D051381:Rats; D058428:Silica Gel; D012822:Silicon Dioxide / Q000737:chemistry; D013997:Time Factors; D014407:Tumor Cells, Cultured
D010761:Phosphorus Radioisotopes; D058428:Silica Gel; D012822:Silicon Dioxide; D002772:Cholera Toxin; D000242:Cyclic AMP; D037342:Pertussis Toxin; D000262:Adenylyl Cyclases; D007545:Isoproterenol