Overview

  • Product name

    Universal Kinase Assay Kit (Fluorometric)
  • Detection method

    Fluorescent
  • Sample type

    Cell Lysate, Purified protein
  • Assay type

    Enzyme activity
  • Sensitivity

    < 0.3 µM
  • Range

    1 µM - 300 µM
  • Species reactivity

    Reacts with: Mammals, Other species
  • Product overview

    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.


     

  • Notes

    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.

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Store 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
  • Research areas

Images

  • 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 

Protocols

References

This product has been referenced in:

  • 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). Read more (PubMed: 30799198) »
  • Danelishvili L  et al. Mycobacterium tuberculosis alters the metalloprotease activity of the COP9 signalosome. MBio 5:N/A (2014). Functional Studies ; Human . Read more (PubMed: 25139900) »
See all 2 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Abreviews
I have tested the Universal Kinase Assay Kit with a member of the Arabidopsis GSK3 kinases, ASKalpha. In general, the kit is user friendly, the provided instructions are easy to follow. The only drawback is that any other tested kinase buffers produced high background, except the buffer which was provided with the kit. In the kinase reactions, as a positive control I used hexokinase, a commercially available kinase. I could detect significant changes in ADP levels in the kinase reactions, so the kit was working fine.

Abcam user community

Verified customer

Submitted Aug 13 2019

Answer

1. This product IS designed to exclusively detect ADP formation. 2. Residual ATP in sample WILL NOT effect the read. 3. To deal with AMP formation, the enzyme coupled reaction in the kit by using ADP is much faster than the formation of AMP.

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