The Adapta® Universal Kinase Assay Kit is a homogenous, fluorescence-based immunoassay for the detection of ADP.  Adapta® Assays utilize Transcreener® HTS Assay Platform technology (patent no 7332278) under license from Bellbrook Labs, LLC. Some of the benefits of this assay format include:
  • High Z’-factors at low percent conversion of ATP to ADP
  • One assay suitable for all kinases, including difficult to assay targets such as lipid kinases
  • Best in class Alexa Fluor® dyes enable a superior red-shifted TR-FRET assay
  • Reduced interference from compound autofluorescence or light scattering from precipitated compounds
  • Complete validated assay system available including substrate and specialized assay buffer


A growing list of Invitrogen kinases have been validated for use with the Adapta® Universal Kinase Assay Kit and corresponding substrates. Please view our Adapta® Assay Reactivity Table to see if your kinase has been validated with the Adapta® Assay.

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Assay principle

The principle of the Adapta® Universal Kinase Assay is shown in Figure 1. The assay itself can be divided into two phases: a kinase reaction phase, and an ADP detection phase. In the kinase reaction phase, all kinase reaction components are added to the well and the reaction is allowed to incubate (typically 60 minutes). After the reaction, a detection solution of Europium-labeled anti-ADP antibody, an Alexa Fluor® 647 labeled ADP tracer, and EDTA (to stop the kinase reaction) is added to the assay well. ADP formed by the kinase reaction (in the absence of an inhibitor) will displace the Alexa Fluor® 647 labeled ADP tracer from the antibody, resulting in a TR-FRET signal decrease. In the presence of an inhibitor, the amount of ADP formed by the kinase reaction is reduced, and the resulting intact antibody-tracer interaction results in a high TR-FRET signal.

Image showcasing the fundamental principle of the Adapta® Universal Kinase Assay, a versatile assay method for analyzing and measuring kinase activity

Figure 1. Schematic principle of the Adapta® Universal Kinase Assay

Assay sensitivity

The Adapta® Universal Kinase Assay measures kinase activity by detecting ADP formation. Most of the signal change occurs in the first 10-20% conversion of ATP to ADP (Figure 2). This is in sharp contrast to kinase assays that measure ATP depletion, in which 20% conversion of ATP to ADP results in only a 20% signal change. As a result, the Adapta® Universal Kinase Assays produces high Z’-factors at low % conversions and is ideally suited for use with kinases with low activity since less ADP has to be formed to achieve an optimal assay window.

Image showcasing the relationship between assay signal and the percentage of ATP conversion to ADP
 Figure 2—Assay signal as a function of percent conversion of ATP to ADP. A representative ATP-ADP titration curve highlights the large change in assay signal at only 20% conversion of ATP to ADP. This makes the Adapta® assay ideally suited for use with kinases with low activity since less ADP has to be formed to achieve an optimal assay window.

Substrate availability

In addition to the Adapta® Universal Kinase Assay Kit, we offer a selection of lipid and peptide based substrates for use with the Adapta® Assay.  Both the lipid and peptide substrates were designed and optimized with assay performance in mind.

Our lipid substrates were designed for optimal performance in the Adapta® Universal Kinase Assay with our PI3 kinases.

  • No re-hydrations, sonication, or extrusion needed, the lipid substrates are ready for immediate use in your kinase assay
  • Formulated for optimal performance with PI3 kinases
  • Get reliable, reproducible performance each and every time, even after multiple freeze-thaw cycles

Since lipid kinases are often highly sensitive to reaction conditions, such as the type and concentration of detergents and salts, we have optimized buffers for each PI3 kinase in our collection.

A list of substrates and buffers currently available from Invitrogen for use in the Adapta® Universal Kinase Assay, can be found below.

Adapta® Assay reactivity table

The following is a list of kinases which have been validated using the Adapta® Universal Assay Kit. Please check back often as we are continuously validating additional kinases.

The table also includes the recommended substrate and buffer to run the assay as outlined in the assay conditions section. 5X Kinase Buffer A is included with the purchase of the Adapta® Assay Kit.

The Adatpa® Reactivity table is currently being updated – to obtain a list of kinases which have been validated using the Adapta® Universal Assay Kit or which reagents to use with your kinase, please submit an online inquiry.

KinaseSubstrateBufferLiterature
CAMK1ZIPtide5X Kinase Buffer APV4391 CAMK1 Adapta Assay Validation1
CDK7/cyclin H/MNAT1CDK7/9tide5X Kinase Buffer APV3868 Cdk7 Adapta Assay Validation1
CDK9/cyclin T1CDK7/9tide5X Kinase Buffer APV4131 Cdk9 Adapta Assay Validation1
DAPK1ZIPtide5X Kinase Buffer APV3969 DAPK1 Adapta Assay Validation1
LRRK2ERM (LRRKtide)5X Kinase Buffer SPV4873 LRRK2 Adapta Assay Validation1
LRRK2 G2019SERM (LRRKtide)5X Kinase Buffer SPV4881 LRRK2 G2019S Adapta Assay Validation1
PIK3C3PI:PS Substrate5X Kinase Buffer QPV5126 PIK3C3 (hVPS34) Adapta Validation1
PIK3CA/PIK3R1PIP2:PS Substrate5X Kinase Buffer RPV4788 PIK3CA_PIK3R1 Adapta Assay Validation
PIK3CGPIP2:PS Substrate5X Kinase Buffer RPV4786 PIK3CG Adapta Assay Validation
SPHK1Sphingosine Kinase Substrate5X Kinase Buffer ASPHK1_Adapta
SPHK2Sphingosine Kinase Substrate5X Kinase Buffer APV5216 SPHK2 Adapta Assay Validation
PIK3CD/PIK3R1PIP2:PS Substrate5X Kinase Buffer RSubmit an online inquiry
PI4KBPI:PS Substrate5X Kinase Buffer TPV5277 PI4KB Adapta Assay Validation
PIK3C2BPI Substrate5X Kinase Buffer RPV5374 PIK3C2B Adapta Assay Validation1
PIK3C2API Substrate5X Kinase Buffer RPV5586 PIK3C2A Adapta Validation1
PI4KAPI:PS Substrate5X Kinase Buffer TPV5689 PI4KA (PI4K alpha) Adapta Validation
MKNK2RS peptide5X Kinase Buffer APV5607 MKNK2 (MNK2) Adapta Validation
GSG2Histone H3 Peptide (1-20)5X Kinase Buffer APV5708 GSG2 Adapta Assay validation
PDK4PDH peptide5X Kinase Buffer APV5710 PDK4 Adapta Assay validation
MKNK1RS peptide5X Kinase Buffer AMKNK1 _MNK1_ Adapta

Instrument settings

Most instruments, settings, and filters that work with other europium-based TR-FRET assay systems will perform well with the Adapta® Universal Kinase Assay. As with other TR-FRET systems, the europium donor is excited using a 340-nm excitation filter with a 30-nm bandpass. Energy transfer to the Alexa Fluor® 647 tracer is measured using a filter centered at 665 nm with a 10 nm bandpass. This signal is then referenced (or “ratioed”) to the emission from europium peak, using a 615 nm, 10-nm bandpass filter (See Figure 3). The “emission ratio” is calculated as the 665 nm signal divided by the 615 nm signal.

For assistance with determining if your instrument is suitable to read the Adapta® Universal Kinase Assay, or for assistance with instrument setup, please call us at 1 760 603 7200 (select option 3, then enter extension 40266).

Graph showing the excitation and emission spectra of the Adapta® Universal Kinase Assay, indicating the wavelengths of light used for excitation and detection of fluorescence

Figure 3. Excitation and emission spectra of the Adapta® Universal Kinase Assay
The emission spectrum of the europium donor (in black) overlaps with the excitation spectrum for the Alexa Fluor® 647 ADP tracer (dashed red line). Following energy transfer, the emission from the Alexa Fluor® 647 ADP tracer (solid red line) is detected with a filter centered at 665 nm with a 10 nm bandpass. This signal is then referenced (or ‘ratioed’) to the emission of the europium signal using a filter centered at 615 nm with a 10 nm bandpass.

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