The ONLY Kinase Assay Method
Available with Direct Detection of ADP

Transcreener ADP2 Kinase Assay Kit

Transcreener ADP Kinase Assay Kit directly measures ADP produced by kinases. These ADP measurements allow researchers to effectively determine the activity of the kinase enzyme. The assay provides a powerful tool to screen compound libraries for kinase modulators to help find new therapies for disease.

The kit comes complete with the detection reagents required to measure activity. Kinase enzyme is not included in the inhibitor screening assay kit. Please contact us for questions related to acquiring the enzyme. This assay is designed to be used with purified enzyme preparations. Transcreener assays are not validated with cell lysates, blood, serum, or other biological samples.

Check out these success stories using Transcreener's ADP Kinase Assay Kit:

  • Eli Lilly used Transcreener to discover and patent Aurora A Kinase Inhibitors for cancer therapeutic development.
  • Researchers at the University of Wisconsin - Milwaukee used the ADP assay to measure helicase activity in an effort to find small molecule inhibitors as a therapeutic for hepatitis C.
  • Pfizer used the assay to find Acetyl-CoA Carboxylase Inhibitors as potential treatments for a variety of indications including diabetes and metabolic syndrome.

How Does This ADP Assay Kit Work?

The Transcreener Kinase Assay Kit determines kinase enzyme activity by directly measuring the ADP formed by the enzyme using the Transcreener ADP2 Assay. By detecting ADP output, the assay provides a universal method to assess the activity of any ADP-producing enzyme in real-time.

Transcreener Kinase Assay technology uses a simple but highly effective method that consists of an antibody selective to ADP over ATP and a far-red fluorescent tracer. ADP produced in the reaction competes with the tracer changing the fluorescent properties and providing fluorescent readout.

The kinase assay kit is available with FP, FI, and TR-FRET. It is a simple mix-and-read format. Perform your enzyme reaction, add the detection reagent, and measure. The simplicity of the system yields robust results that also makes it extremely amiable to HTS.

Example Applications

Direct Detection of ADP to Measure Enzymatic Activity

Fluorescent Polarization (FP)

ADP FP Kinase Assay Kit

The workhorse. Used in many large screens. Best deck and signal stability.

Fluorescent Intensity (FI)

ADP FI Kinase Assay Kit

Positive FI signal. Compatible with simple fluorescence plate readers. Faster read time than FP or TR-FRET.

Time-Resolved Förster Resonance Energy Transfer (TR-FRET)

ADP TR-FRET Kinase Assay Kit

For customers who prefer TR-FRET detection. Uses the same filter set as HTRF®.

Applications

  • Measure Enzymatic Activity of Kinases
  • Screen Compound Libraries for Kinase Modulators
  • Quantify Inhibitor Potency
  • Inhibitor Selectivity Profiling
  • Measure Drug-Target Residence Time

Features

  • Direct detection of unlabeled ADP
  • Easy to use, homogenous, one-step format
  • Robust Assay Z' > 0.7 under initial velocity conditions
  • Far-red fluorescent readouts minimize compound interference
  • A safe, non-radioactive method
  • Use with ATP concentrations from 0.1 to 1000 µM
  • Available in FP, FI, or TR-FRET readouts
  • Can be used in continuous or endpoint mode

Easy-to-Use, Mix-and-Read, HTS Ready Assay

Run your enzyme reaction, add Transcreener reagents, and read your plates. The kinase assay kit is compatible with 96, 384, and 1536-well formats. 

Mix-and-Read Simplicity

Universal – Use with Virtually Any Kinase

Overcome the need for time-consuming, one-off development for individual members within an enzyme family by using a single set of reagents that detect the invariant product – ADP. The assay can be used with virtually any substrate including peptide substrates, protein substrates, and lipid substrates.

Example enzyme targets include:

  • Protein kinases
  • Lipid kinases
  • Carbohydrate kinases
  • ATPases
  • Chaperonins (Hsp90, etc.)
  • Nucleotidases
  • Carboxylases
  • Helicases

Rock-Solid Reagent and Signal Stability Ensures High-Quality Data and Simplifies Automation

ADP Assay Reagent And Signal Stability Data

The Transcreener ADP2 antibody and tracers are stable at room temperature for a minimum of 8 hours. The outstanding reagent and signal stability make Transcreener ADP2 Kinase Assays very reliable and easy to use in an automated HTS environment.

Kinase Assay Services

Interested in moving your program forward, but don't want to bring an assay in-house? Our scientists can help! BellBrook scientists will use their extensive biochemistry and enzymology expertise to work with you and accelerate your kinase program.

Assay Development Services

Lead Discovery Services Include:

  • Inhibitor Screening - To identify or confirm activity with the target.
  • Inhibitor Potency Profiling - Dose-response with target and/or related proteins. Fast IC50 results.
  • Residence Time Measurements - Determination of koff using 'jump dilution' enzymatic assay method.
  • Mechanism of Action Studies - Kinetic analysis to define the mode of inhibition.

Contact Us to Learn More

Please fill out the form below. We will respond quickly to get the conversation moving and learn how we can help. We keep things discrete, confidential, and professional. 

Far Red FP, FI & TR-FRET Readouts Validated on Major Multimode Readers

SupplierInstrumentFP AssaysFI AssaysTR-FRET Assays
TriStar2S LB 942In ReviewClick Me ArrowValidatedClick Me ArrowValidated
Tristar 5In ReviewClick Me ArrowValidatedIn Review
Mithras2 LB 943In ReviewClick Me ArrowValidatedClick Me ArrowValidated
CytationTM 5ValidatedValidatedValidated
CytationTM 3ValidatedValidatedValidated
CytationTM 1ValidatedValidatedValidated
SynergyTM H1ValidatedValidatedValidated
SynergyTM 2/H4/4Click Me ArrowValidatedClick Me ArrowValidatedClick Me ArrowValidated
SynergyTM HTXNot CapableValidatedNot Capable
SynergyTM Neo 2ValidatedValidatedValidated
POLARstar® OmegaClick Me ArrowValidatedClick Me ArrowValidatedClick Me ArrowValidated
FLUOstar® OmegaNot CapableClick Me ArrowValidatedClick Me ArrowValidated
PHERAstar® FSXClick Me ArrowValidatedClick Me ArrowValidatedClick Me ArrowValidated
PHERAstar® Plus/FSClick Me ArrowValidatedClick Me ArrowValidatedClick Me ArrowValidated
CLARIOstar® /PlusClick Me ArrowValidatedClick Me ArrowValidatedClick Me ArrowValidated
VANTAstarTMClick Me ArrowValidatedClick Me ArrowValidatedClick Me ArrowValidated
SenseIn ReviewClick Me ArrowValidatedIn Review
Analyst® GT/HTClick Me ArrowValidatedClick Me ArrowValidatedClick Me ArrowValidated
Gemini® XPS/EMNot CapableClick Me ArrowValidatedNot Capable
SpectraMax® M2/M2eNot CapableClick Me ArrowValidatedNot Capable
SpectraMax® M5/M5e/FlexStation® 3Not CapableClick Me ArrowValidatedClick Me ArrowValidated
SpectraMax® ParadigmClick Me ArrowValidatedClick Me ArrowContact UsClick Me ArrowContact Us
SpectraMax® iD3/iD5Click Me ArrowValidatedClick Me ArrowValidatedClick Me ArrowValidated
SpectraMax® i3xClick Me ArrowValidatedIn ReviewIn Review
EnVision®/EnVision® XciteClick Me ArrowValidatedClick Me ArrowValidatedClick Me ArrowValidated
Infinite® M1000/M1000Pro/Safire2TMClick Me ArrowValidatedClick Me ArrowValidatedClick Me ArrowValidated
Infinite® M200Not CapableClick Me ArrowValidatedNot Capable
Infinite® F500Click Me ArrowValidatedClick Me ArrowValidatedClick Me ArrowValidated
Infinite® F200/UltraevolutionClick Me ArrowContact UsClick Me ArrowValidatedClick Me ArrowContact Us
SparkTM 10MClick Me ArrowValidatedClick Me ArrowValidatedClick Me ArrowValidated

In addition to giving you flexibility of three detection modes, we have collaborated with the major suppliers of multimode readers, including Tecan, BMG, and Biotek to determine optimal instrument hardware and software settings for each of the Transcreener ADP2 Assays. Successful validation requires a Z’ of greater than 0.7 for ADP detection under initial velocity conditions (≤10% ATP conversion). Optimal filter sets and instrument settings are summarized in the technical manual for each Transcreener assay, and more detailed information is available in application notes. This ensures that whatever fluorescent detection mode or reader you prefer, you will get robust results with Transcreener reagents.

What's Included

What You Will Need

Related Resources

A Guide to Measuring Drug-Target Residence Times with Biochemical Assays

During drug development initiatives, analysis of drug-target residence times can improve efficacy, increase therapeutic window, and reduce the risk of premature focus on candidate compounds that are likely to have undesirable side effects. This guide provides technical background on concepts and techniques for use of Transcreener biochemical assays to measure drug-target residence times, along with examples and case studies.

Learn more including:

  • The basics of residence time and its importance to drug discovery
  • A comparison of methods used for residence time determination
  • Using the jump-dilution method with Transcreener biochemical assays
  • Data analysis used to accurately determine residence time
  • Examples of measuring residence time for kinases, phosphodiesterases, and glycosyltransferases
A Guide to Measuring Drug-Target Residence Times with Biochemical Assays

Enter your contact info and learn how to determine drug-target residence times and improve your lead discovery with this 12-page guide!

Targeting Kinases Involved in the Innate Immune Response

Clinical trials continue to determine whether therapeutic modulation of kinases can be used to suppress the immune system in autoimmune diseases or stimulate for antiviral and cancer immunotherapy. The discovery and characterization of small molecule modulators have aided these efforts.

The Transcreener ADP² Kinase Assay meets this need as it relies on direct ADP detection to measure the activity of virtually any kinase. The method has been extensively validated for kinase discovery programs since 2007. This article provides five examples of how Transcreener allowed rapid assay development to enable screening and dose-response measurements.

The innate immunity-associated kinases targeted in the five case studies include:

  • AMP-Activated Kinase (AMPK)
  • Janus Kinase 1 and 3 (JAK1 and JAK3)
  • TANK-Binding Kinase 1 (TBK1)
  • Interleukin-1 Receptor-Associated Kinase 4 (IRAK4)
  • IκB Kinase (IKK-β)
Targeting Kinases in Innate Immunity

Enter your contact info and learn more about targeting innate immune-associated kinases in this free 12-page guide.

Kinases As Therapeutic Targets

Kinases transfer a phosphate to a protein. Kinases play important roles in cell proliferation, survival, and migration. They regulate cellular function through their ability to turn protein function on. When mutation or deregulation occurs, kinases have been associated with carcinogenesis and metastases of various types of cancer (Bhullar et al, 2018).

Examples of current kinase inhibitors used in clinical treatments include imatinib and dasatinib. These treatments have seen great success for myeloid leukemia (CML) and gastrointestinal stromal tumors (GIST), showing a significant increase in patient survival. The FDA has now approved several small-molecule kinase inhibitors (Bhullar et al, 2018).

The Transcreener ADP2 Kinase Assay is an excellent tool for researchers examining kinases for therapeutic treatments.

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