Recombinant human ABL1 (E255K) (27–end) was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. ABL1 protooncogene encodes a cytoplasmic and nuclear protein tyrosine kinase that has been implicated in processes of cell differentiation, cell division, cell adhesion and stress response. Activity of ABL protein is negatively regulated by its SH3 domain, and deletion of the SH3 domain turns ABL1 into an oncogene (1). Translocation and head-to-tail fusion of the BCR and ABL1 genes is present in many cases of chronic myelogeneous leukemia (2). The DNA-binding activity of the ubiquitously expressed ABL1 tyrosine kinase is regulated by CDK1-mediated phosphorylation, suggesting a cell cycle function for ABL1.
ADP-Glo™ Kinase Assay is a luminescent kinase assay that measures ADP formed from a kinase reaction; ADP is converted into ATP, which is a substrate in a reaction catalyzed by Ultra-Glo™ Luciferase that produces light. The luminescent signal positively correlates with ADP amount and kinase activity. The assay is well suited for measuring the effects chemical compounds have on the activity of a broad range of purified kinases, making it ideal for both primary screening as well as kinase selectivity profiling. The ADP-Glo™ Kinase Assay can be used to monitor the activity of virtually any ADP-generating enzyme (e.g., kinase or ATPase) using up to 1mM ATP.
Kinase Enzyme System contains:
Kinase: ABL1 (E255K), 10μg (Human, recombinant; amino acids 27–end). MW: ~160kDa. Substrate: Abltide (EAIYAAPFAKKK); derived from the C-terminus of ABL. Other: Reaction Buffer, MnCl2, DTT.
Profile More Compounds In-House: ADP-Glo™ Kinase Assay + Kinase Enzyme System is optimized so that you are up and running in no time. Complete Systems: The Kinase Enzyme Systems include a recombinant kinase enzyme, a substrate appropriate for the enzyme, a reaction buffer, DTT and supplemental reagents as needed. Obtain Reliable Results: The broad dynamic range, the ease of use and better sensitivity obtained with ADP-Glo™ Kinase Assay result in less ambiguous data.
Notes
Kinase Enzyme System manufactured by SignalChem.
Bulk quantities available upon request.
References
1.Barila, D. et al. (1998) An intramolecular SH3-domain interaction regulates c-Abl activity. Nature Genet. 18, 280–2.
2.Goldman, J.M. et al. (2001) Targeting the BCR-ABL tyrosine kinase in chronic myeloid leukemia. New Eng. J. Med. 344, 1084–6.
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