Transcreener® AMP²/GMP² Phosphodiesterase Assay Kits

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Single-Step Phosphodiesterase Assay Kit for Screening and Lead Discovery

Simple mix-and-read format
Ultra-sensitive - Use less enzyme
Use with any AMP or GMP-producing enzyme - Including PDEs
Flexible - Use continuous or end-point assay methods

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Transcreener AMP²/GMP² Phosphodiesterase Assay Kits

Transcreener AMP2/GMP2 Phosphodiesterase Assay directly measures AMP or GMP produced by a PDE reaction, making it a powerful phosphodiesterase assay for real product detection. This assay allows detection of any AMP or GMP producing enzyme (e.g. ligases, synthetases, phosphodiesterase) using any precursor substrate, including cAMP, cGMP, ATP, or NAD.

It is the only mix-and-read AMP assay kit solution for direct detection of unlabeled AMP (or GMP) produced by PDEswithout coupling enzymes, delivering a truly universal AMP/GMP readout.

The kit comes complete with all the necessary detection reagents required to measure activity.

Note: It does not include enzyme. 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.

How Does the Assay Work?

The Transcreener AMP²/GMP² Assay determines enzyme activity by directly measuring AMP or GMP formed by the enzyme. By detecting AMP or GMP, the assay provides a universal method to assess the PDE activity of any AMP/GMP-producing enzyme in real-time.

Transcreener technology uses a simple but highly effective method that consists of an antibody selective to AMP or GMP and a far-red fluorescent tracer. AMP/GMP produced in the reaction competes with the tracer changing the fluorescent properties and providing fluorescent readout.

The assay is available with FP or TR-FRET in 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 make it extremely amiable to high throughput screening.

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

Profiling PDEs with the Transcreener AMP²/GMP² Assay

Specifications

What’s Included

FP
	1K		10K
Component	Part #	Qty	Part #	Qty
AMP²/GMP²AlexaFluor® 633 Tracer, 800 nM	2097	100 µL	2101	1 mL
AMP²/GMP² Antibody, 1.26 mg/mL	2098	55 µL	2102	550 µL
1M Tris (pH 7.5)	2132	250 µL	2133	2.5 mL
5 mM AMP	2099	200 µL	2103	2 mL
5 mM GMP	2100	200 µL	2104	2 mL

TR-FRET
	1K		10K
Component	Part #	Qty	Part #	Qty
AMP²/GMP² Hilyte 647 Tracer, 10 µM	2157	200 µL	2159	2 x 1 mL
AMP²/GMP² Antibody Tb, 800 nM	2158	100 µL	2160	1 mL
1M Tris (pH 7.5)	2132	250 µL	2133	2.5 mL
5 mM AMP	2099	200 µL	2103	2 mL
5 mM GMP	2100	200 µL	2104	2 mL

What You Will Need

Component	Notes
Enzyme	The assays are designed for use with purified enzymes. Use any enzyme that produces AMP, GMP, or CMP.
Enzyme Buffer	Enzyme buffer components optimized for your target. See the technical manual for potential interfering components.
ATP, cAMP, cGMP	Use the nucleotide required for your enzyme. It is possible to purchase some nucleotides direct from BellBrook Labs.
Assay Plates	Assay Plates Can Be Purchased Separately From BellBrook Labs. FP - An entirely black plate with a non-binding surface is required. We suggest Corning 384-well plates (Cat. #4514) or 1536-well plates (Cat. #3728). TR-FRET - An entirely white plate with a non-binding surface is required. We suggest Corning 384-well plates (Cat. #4513) or 1536-well plates (Cat. #3729).
Stop Buffer	Some PDEs and Ubiquitin Ligases have inhibited activity with Stop Buffer B (Cat. #2027-1 mL, Cat. #2032 10 mL) which quenches MgCl₂.This allows the assay to be run in endpoint mode.
Plate Reader	A multi-detection microplate reader configured to measure the output of the AMP/GMP tracer is required.
Liquid Handling Devices	Use liquid handling devices that can accurately dispense sub-microliter volumes into 384-well plates.
Ultrapure Water	Some deionized water systems are contaminated with nucleases that can degrade both nucleotide substrates and products, reducing assay performance. Careful handling and use of ultrapure water eliminates this potential problem.

Applications for the Phosphodiesterase Assay Kit

Measure Enzymatic Activity of Phosphodiesterase
Screen Compound Libraries for Phosphodiesterase Inhibitors
Quantify Inhibitor Potency

Inhibitor Selectivity Profiling
Measure Drug-Target Residence Time

Features

Direct detection of unlabeled AMP or GMP
Easy to use, homogenous, one-step format
Robust Assay Z’ > 0.7 under initial velocity reactions
Far-red fluorescent readouts minimizes compound interference
A safe, non-radioactive assay technique
Wide substrate concentration range: 1 μM to 1000 µM

One Assay … Hundreds of Targets

A universal AMP/GMP readout beyond PDEs.
Along with PDEs the kit can be used with:

Ligases: Ub and SUMO Ligases, DNA and RNA Ligases
Synthetases: Acyl CoA Synthetase, Amino-Acyl tRNA Synthetases, NAD Synthetase
Phosphodiesterases: cAMP, cGMP
Sialyltransferases: (CMP detection)
Protein deAMPylase
Ectonucleotidases such as CD39 and ENPP1
Exoribonucleases such as XRN1

Sensitive Assay with Large Window

See real activity changes across substrate and inhibitor ranges

Measure the Potency of Inhibitors for a Variety of Targets

Fast IC₅₀ and selectivity reads without assay re-engineering

Because the readout is tied to AMP/GMP products, not the substrate, your assay setup remains consistent across different PDE families and related enzyme targets, accelerating potency profiling and selectivity panels

Potency of Inhibitors PDE41A and UbE1 Dose Response Curves

Easy-to-Use, Mix-and-Read, HTS-Ready GMP/AMP Assay

Designed for automation and scale

Run your enzyme reaction
Add Transcreener reagents
Read your plates

The assay is compatible with 96, 384, and 1536-well formats, making this phosphodiesterase assay kit ideal for HTS and SAR workflows.

Transcreener Mix and Read Assay Schematic

Phosphodiesterase 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 PDE program.

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 k_off using ‘jump dilution’ enzymatic assay method.
Mechanism of Action Studies – Kinetic analysis to define the mode of inhibition.

Scientist pipetting into a microplate beside a microscope.

Contact Us for Services

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.

PDE’s As Enzyme Drug Targets

Phosphodiesterase’s (PDE’s) are enzymes that catalyze the hydrolysis of cAMP or cGMP to form AMP or GMP. cAMP and cGMP are classic cyclic monophosphate messengers that play important roles in a variety of biochemical processes. These cyclic nucleotides regulate major signaling pathways across tissues, and PDEs are organized into 11 families based on substrate preference, structure, and distribution. Differences in cyclic nucleotide phosphodiesterase activity across families help determine how quickly signals are turned off in specific tissues.

Because PDEs control the balance of cyclic GMP AMP signaling, they have excellent promise in a variety of therapeutic areas, including heart disease, dementia, infection, depression, and asthma. New inhibitors can provide better therapeutic potential and can limit off-target effects more effectively.

Cyclic nucleotide phosphodiesterases are proven drug targets. Most are a focus of inhibition due to their functional properties. Since PDEs degrade cAMP or cGMP into inactive monophosphates, including monophosphate cAMP, inhibitors can prolong the effects of the cyclic nucleotide in the signaling cascade within the tissues where the individual enzyme is most likely to reside.

Substrate Specificity

Nucleotide	PDE
cAMP	1, 2, 3, 10, 11
cGMP	5, 6, 9
cAMP & cGMP	1, 2, 3, 10, 11

The Transcreener enables researchers to scour vast compound libraries for potent, yet selective inhibitors for unique PDEs. You can also complete a follow-up SAR to improve upon initial hits and increase the chance of success.

Related Resources

Resources

Find the right resources to help you get to your solution faster.

FAQ

Protocols

Publications

Instrument Compatibility

Application Notes

Posters

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.

“Some CROs reserve their best teams for high profile customers. If you’re not right there with them you get less experienced teams… BellBrook understands enzyme kinetics, so it made that part of the project so much easier. The larger CRO didn’t seem to have the expertise we needed in this area.”

- BioPharma Lead Discovery Program Manager

“The ADP assay is a great choice for measuring activity for any ADP-generating enzyme in both real time and endpoint formats. The universal detection system has been very useful for more challenging targets.”

- Pharmaceutical Research Scientist

“Excellent blend of simplicity & good science. Excellent technical support”

- Director Pharma Drug Discovery Center

“The assay itself has good reproducibility and provides a clear readout on our enzyme activity. The most important piece, however, is the level of attention to our project and rapid turnaround time. The staff at BellBrook knows what they are doing.”

- Biotech Program Manager

“The Transcreener AMP/GMP Assay is extremely straightforward to use and is very sensitive. In a UV-based enzyme assay I needed 1 µM enzyme to see a decent signal; however using the Transcreener AMP assay I only need 10 nM. Also, BellBrook Labs have been very helpful with the assay development. Their representative has been very willing to quickly help me get my assay up and running.”

- University Research Scientist

“Through the process of developing and validating an assay for HTS applications, I generated a very nice data package of robust high quality data.”

- Biotech Research Scientist

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Frequently Asked Questions

What makes the Transcreener AMP²/GMP² Phosphodiesterase Assay Kit different from other PDE assays?

This kit directly detects AMP or GMP—the products of phosphodiesterase activity—using highly selective antibodies in a homogeneous mix-and-read format. It bypasses coupling enzymes and separation steps, reducing interference and streamlining workflows.

Can this assay detect enzymatic activity besides PDEs?

Yes. Although designed for phosphodiesterases (cAMP/cGMP hydrolysis), this assay works universally for any enzyme that generates AMP or GMP, including ligases, synthetases, and ubiquitination-related enzymes.

Which detection modes are supported?

Both fluorescence polarization (FP) and TR‑FRET readouts are available, using far-red tracers to minimize compound interference and deliver high-quality data.

Is the kit suitable for high-throughput screening (HTS)?

Absolutely. The kit is validated for 96-, 384-, and 1536-well formats. It delivers excellent signal sensitivity (with Z’ ≥ 0.7 at approximately 10% substrate conversion) even at low native substrate concentrations (1–1000 µM).

How do I get started with the Transcreener AMP²/GMP² PDE Assay Kit?

You can request a sample, download the technical manual, or reach out to our scientific team for guidance on assay setup, plate reader compatibility, or custom screening workflows.

BellBrook Labs
1232 Fourier Drive, Suite 115
Madison, Wisconsin 53717 USA
(608) 443-2400

info@bellbrooklabs.com

Single-Step Phosphodiesterase Assay Kit for Screening and Lead Discovery

Corning® Assay Plates