Promega Corporation

ADP-Glo™ Kinase Assay

ADP-Glo Kinase Assay Work Flow

8051MB

The New Gold Standard in Kinase Assays

  • Sensitive down to 20nM ADP: Less enzyme needed per reaction thus reducing costs; easily miniaturizable.
  • Strong signal at low % conversion of substrate: Obtain meaningful results under initial reaction conditions; also means less enzyme is required.
  • Simple to use: No time wasted optimizing or learning to perform assays; antibody-free.
  • Works with broad range of kinases: Assay more kinases in-house— reduces dependency on outsourcing.
  • Accurate IC50s— results comparable to radio-labeled assays: Allows researchers to switch to a safer method with the peace of mind that their results are comparable to the "gold standard".

Sensitivity and linearity of ADP-Glo Kinase Assay

8052MA-mod

Each graph represents the total concentration of nucleotide (ATP + ADP) in the reaction.

Strong signal to background at low % conversion of substrate

Enzyme SB5* % ATP to ADP** conversion at SB5
EGFR 0.3ng 2.2
PDGFRα 21ng 4.7
VEGFR2 (KDR) 1ng 4.2
DNA-PK 0.4ng 2.7
AKT2 7ng 4.3
IKKβ 4.5ng 2.9
MAPK (ERK2) 1ng 5.6
PKA 0.013 units 0.35
Hexoklnase 0.1 units 4.6
PI3 Kinase γ 0.15ng 2
Sphingosine Kinase 1 0.8ng 2.9
MDR1 (P-gp) 25μg (SB20) 50
Na+/K+ ATPase 1ng 3

* Amount of enzyme required to produce a signal to background ratio of 5 using ADP-Glo.
** Enzyme reactions were performed at different ATP Concentrations.

Works with broad range of kinases

8193MA-8196MA-mod

Accurate IC50 Values

8197MA

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