PMC:6365940 / 12728-13596 JSONTXT

{"target":"http://pubannotation.org/docs/sourcedb/PMC/sourceid/6365940","sourcedb":"PMC","sourceid":"6365940","source_url":"https://www.ncbi.nlm.nih.gov/pmc/6365940","text":"2.11 Measurement of PI3K activity\nPI3K activity was detected by a PI3K activity ELISA kit (Echelon Biosciences Inc., Salt Lake City, UT, USA) according to the manufacturer's instructions. Briefly, PI3K protein (Sigma-Aldrich, St. Louis, USA) was reacted with PI(4,5)P2 as a substrate in a reaction buffer without and with 80 nM of digoxin at 30 ℃ for 2 h. After termination of the reaction, a primary PI(3,4,5)P3 detector was added to the reaction mixture and incubated for 60 min at room temperature. The mixture was transferred to a PI3K ELISA plate, and 60 min later a second detector was added and further incubated at room temperature for 30 min. A tetramethylbenzidine solution was added to the final reaction mixture, and absorbance were measured at 450 nm using a microplate reader. PI(3,4,5)P3 production (pmol/2 h) was regarded as an index of PI3K activity.","divisions":[{"label":"label","span":{"begin":0,"end":4}},{"label":"title","span":{"begin":6,"end":34}}],"tracks":[]}