Transcreener® ADP² Kinase Assay

The ONLY Kinase Assay Method
Available with Direct Detection of ADP

Transcreener Kinase Assay Principle

Transcreener assays are competitive immunoassays in a mix-and-read format. Displacement of tracer from the ADP antibody causes a change in its fluorescence properties. Transcreener is the simplest, most direct method for measuring ADP formed during a kinase reaction. Since the introduction of the first product in 2006, Transcreener has been used in millions of wells for HTS and lead discovery.

Biochemical kinase assay kits are available in far-red FP, FI, and TR-FRET detection modes to ensure optimal performance with your plate reader.

ADP FP Kinase Assay

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

ADP FI Kinase Assay

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

ADP TR-FRET ATPase Assay

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

Transcreener is 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

Learn how Eli Lilly used Transcreener to discovery and patent Aurora A Kinase Inhibitors for cancer therapeutic development.

See how 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.

Discover how Pfizer used the assay to find Acetyl-CoA Carboxylase Inhibitors as potential treatments for a variety of indications including diabetes and metabolic syndrome.

Applications

  • Screen for small molecule inhibitors
  • Determine enzymatic activity
  • Profile inhibitor potency/determine IC50 values
  • Measure residence time/off-rate/dissociation of lead molecules with target
  • Inhibitor selectivity profiling
  • Mechanism of action studies

Features

  • Simple homogenous biochemical assay
  • Direct detection of ADP eliminates laborious coupling steps
  • Non-radioactive, safe assay method
  • HTS Ready – Use in 96-well, 384-well, or 1536-well formats
  • Choose from Fluorescent Polarization (FP), Fluorescent Intensity (FI), or time-resolved Förster-resonance-energy-transfer (TR-FRET) formats
  • Sensitive, robust detection of ADP. This means up to 10X less enzyme, a significant cost reduction on large screens.
  • Use with ATP concentrations from 0.1 to 1000 µM
  • Far-red tracer further minimizes interference from fluorescent compounds and light scattering

Direct ADP Detection Minimizes Interference

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Learn why direct ADP detection is the best HTS assay method for Kinases and ATPases

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Transcreener ADP2 Kinase Assay Kits rely on direct detection of ADP using a proprietary antibody that binds with exquisite specificity and affinity to ADP, with negligible cross-reactivity to ATP. This exquisite selectivity allows direct detection of ADP in the presence of an excess of ATP, a requirement for kinase activity assays run under initial velocity conditions (e.g., ≤10% ATP consumption). The high affinity of the ADP antibody means that the assay has the sensitivity for robust detection of ADP down to 10 nM with Z’ values of 0.7 or greater, so you can use less of your precious enzyme.

Other ADP detection methods rely on complex assay schemes that use three or more enzymatic steps to convert ADP into a detectable signal. Each of the enzymes in the detection cascade is a potential target for the compounds being screened, which increases the risk of false positives or of missing a hit, and requires additional wells for counter-screening.

Mix-And-Read Format Can Be Used in Endpoint or Continuous Mode

Transcreener ADP2 Assays all use a homogeneous, mix and read assay format, which simplifies automation and provides maximal flexibility for assay protocols. Endpoint assays are typically used for HTS, and continuous assays are used for assay development or in determining residence times for kinase inhibitors.

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!

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

The Transcreener ADP2 antibody and tracers are stable at room temperature for a minimum of 8 hours. This ensures that data quality does not suffer in situations where reagents are stored at room temperature prior to dispensing or if plates are not read immediately. The outstanding reagent and signal stability make Transcreener ADP2 Kinase Assays very reliable and easy to use in an automated HTS environment.

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

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.

Transcreener ADP² Kinase Assay Kits

* For custom or bulk orders (over 100,000 wells) please contact us (info@bellbrooklabs.com) for a quote.

Frequently Asked Questions

Related Resources

In addition to our ADP assays, Bellbrook Labs also offers the following products and services to aid in the drug discovery process:

Methyltransferase Assays

BellBrook Labs
5500 Nobel Drive, Suite 230
Madison, Wisconsin 53711 USA
(608) 443-2400

info@bellbrooklabs.com

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