Key features and details
- Assay type: Enzyme activity
- Detection method: Fluorescent
- Platform: Microplate reader
- Sample type: Cell Lysate, Purified protein
- Sensitivity: 0.3 µM
Product nameUniversal Kinase Assay Kit (Fluorometric)
Sample typeCell Lysate, Purified protein
Assay typeEnzyme activity
Sensitivity< 0.3 µM
Range1 µM - 300 µM
Species reactivityReacts with: Mammals, Other species
Universal Kinase Assay Kit (Fluorometric) (ab138879) is based on monitoring ADP formation, which is directly proportional to enzyme phosphotransferase activity and is measured fluorimetrically. This enzyme-coupled kit provides a fast, simple, and homogeneous assay to measure kinase activities. It is a non-radioactive and no wash method to detect the amount of ADP produced from enzyme reaction. Its characteristics of high sensitivity (<0.3 μM ADP) and broad ATP tolerance (1-300 μM) make it an ideal kit for determining kinase Michaelis-Menten kinetics and for screening and identifying kinase inhibitors. The assay can be performed in a convenient 96-well or 384-well microtiter plate format and easily adapted to automation without a separation step.
Protein kinases are the enzymes that transfer a phosphate group from a phosphate donor to an acceptor amino acid in a substrate protein. Kinases are of great interest to researchers involved in drug discovery. Most of the commercial protein kinase assay kits are based on monitoring either the phosphopeptide formation or the ATP depletion. For the kinase assay kits that are based on the detection of phosphopeptides, one has to spend time and efforts to identify an optimized peptide substrate while the ATP depletion method suffers various interferences due to the use of luciferase that are inhibited or activated by various biological compounds.
Storage instructionsStore at -80°C. Please refer to protocols.
Components 250 tests ADP Assay Buffer 1 x 10ml ADP Sensor Buffer 1 x 5ml ADP Sensor I (Light-sensitive) 1 vial ADP Sensor II 1 x 2.5ml ADP Standard 1 vial DMSO 1 x 100µl
ADP dose response was measured with ab138879 in a solid black 384-well plate using a fluorescence microplate reader. As low as 0.3 µM ADP can be detected with 15, 30 minutes and 1 hour incubation (Z’ factor =0.65).
The Universal Kinase Assay Kit is based on the monitoring of ADP formation, which is directly proportional to enzyme phoshphotransferase activity and is measured fluorimetricaly
ab138879 has been referenced in 5 publications.
- Terekhov SS et al. A kinase bioscavenger provides antibiotic resistance by extremely tight substrate binding. Sci Adv 6:eaaz9861 (2020). PubMed: 32637600
- Bresch AM et al. The PP1 regulator PPP1R2 coordinately regulates AURKA and PP1 to control centrosome phosphorylation and maintain central spindle architecture. BMC Mol Cell Biol 21:84 (2020). PubMed: 33238888
- Sun K et al. Oxidized ATM-mediated glycolysis enhancement in breast cancer-associated fibroblasts contributes to tumor invasion through lactate as metabolic coupling. EBioMedicine 41:370-383 (2019). PubMed: 30799198
- Tsang CK et al. SOD1 Phosphorylation by mTORC1 Couples Nutrient Sensing and Redox Regulation. Mol Cell 70:502-515.e8 (2018). PubMed: 29727620
- Danelishvili L et al. Mycobacterium tuberculosis alters the metalloprotease activity of the COP9 signalosome. MBio 5:N/A (2014). Functional Studies ; Human . PubMed: 25139900