Kinase Assay (Method B): The kinase activity of the bi-functional enzyme is readily quantified based on the production of ADP and F-2,6-P2 from ATP an...

ID: ALA3887842

Type: Binding

Description: Kinase Assay (Method B): The kinase activity of the bi-functional enzyme is readily quantified based on the production of ADP and F-2,6-P2 from ATP and F6P. The ADP is detected with a kit, ADP-Glo Kinase Assay(a-e), from Promega. The assay is a luminescent ADP detection assay that measures kinase activity by quantifying the amount of ADP produced during the kinase reaction. The assay is performed in two steps; first, after the kinase reaction, an equal volume of ADP-Glo Reagent is added to terminate the kinase reaction and deplete the remaining ATP. Second, the Kinase Detection Reagent is added to simultaneously convert ADP to ATP and allow the newly synthesized ATP to be measured using a luciferase/luciferine reaction. The light generated is measured using a luminescence counter (1450 MicroBeta TriLux).The assay was performed in white 384-well microtiter plates (OptiPlate, 6007299, PerkinElmer) by consecutive additions of 0.1 uL of a test compound solution.

Format: BAO_0000357

Target:  6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 4(ALA3721311)

Document:  ALA3886470

BindingDB Source:  7548