The MicroMolar Universal Kinase Assay Kit is based on measurement of ADP generated form the kinase reaction. It is used to measure the activities of purified kinases producing 5 mM – 100 mM (micromolar) ADP. The assay is fluorescence-based and can be carried out using regular black or white 96-well or 384-well plates or micro-cuvettes. It is important to avoid using unnecessarily high ATP concentrations in the kinase assays. There are major two reasons for this. One is to allow the assay to detect ATP-competitive inhibitors. The other is to avoid a high ADP background from ADP contamination in the ATP samples. It is recommended to use an ATP concentration in the range of 30 mM to 300 mM (micromolar) depending on the Km of ATP for the kinase.
The kit contains the 10 x reaction buffer (10 x MUK buffer), 100 x MUK reagent 1, 100 x MUK reagent 2 and 10 x MUK dye. The kit reagents are sufficient for 100 assays of purified kinases using a standard black or white 384-well plate.
The kit contains the 10 x reaction buffer (10 x MUK buffer), E. coli thymidylate Kinase, dTMK substrate, 100 x MUK reagent 1, 100 x MUK reagent 2 and 10 x MUK dye. The kit reagents are sufficient for 100 assays of E. coli thymidylate kinase using a standard black or white 384-well plate.
