Profiling PDEs with the Transcreener AMP²/GMP² Phosphodiesterase Assay

Transcreener AMP2/GMP2 Phosphodiesterase Assay Kit

Transcreener AMP2/GMP2 Phosphodiesterase Assay directly measures AMP or GMP produced by phosphodiesterase enzymes. These AMP/GMP measurements allow researchers to effectively determine the activity of the phosphodiesterase enzyme. The assay provides a powerful tool to screen compound libraries for phosphodiesterase modulators to help find new therapies for disease. 

The kit comes complete with the detection reagents required to measure activity. Phosphodiesterase enzyme is not included in the inhibitor screening assay kit. Please contact us for questions related to acquiring the enzyme.

Transcreener AMP2/GMP2 Assay kits allow 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 activity assay method for direct detection of unlabeled AMP or GMP; i.e. without coupling enzymes.

How Does This Phosphodiesterase Assay Work?

The Transcreener AMP2/GMP2 Assay determines phosphodiesterase 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 activity of any AMP/GMP-producing enzyme in real-time. 

Transcreener Phosphodiesterase Assay 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 Phosphodiesterase Assay is available with FP or 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 make it extremely amiable to HTS. 

Universal AMP/GMP Detection for Phosphodiesterases

Fluorescent Polarization (FP)

Transcreener AMP GMP FP Assay Schematic

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

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

Transcreener AMP GMP TR-FRET Assay Schematic

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

Applications

  • 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

Along with PDEs the Phosphodiesterase Assay 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, CD73, and ENPP1

Sensitive Assay with Large Window

Graph of Sensitivity Window of PDE41A

Measure the Potency of Inhibitors for a Variety of Targets

Potency of Inhibitors PDE41A and UbE1 Dose Response Curves

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

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

Transcreener Easy-Mix-and Read Method

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.

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 & TR-FRET Readouts Validated on Major Multimode Readers

Supplier Instrument FP Assays
TR-FRET Assays
bioteklogo TriStar²S LB 942 in review validated
Mithras² LB 943 in review validated
bioteklogo Cytation™ 5 validated validated
Cytation™ 3 validated validated
Cytation™ 1 validated validated
Synergy™ H1 validated validated
Synergy™ 2/H4/4 validated validated
Synergy™ HTX not capable not capable
Synergy™ Neo 2 validated validated
BMGLABTECH Logo POLARstar® Omega validated validated
FLUOstar® Omega not capable validated
PHERAstar® FSX validated validated
PHERAstar® Plus/FS validated validated
CLARIOstar® /Plus validated validated
VANTAstar validated validated
MDS AT logo Analyst® GT/HT validated validated
Gemini® XPS/EM not capable not capable
SpectraMax® M2/M2e not capable not capable
SpectraMax® M5/M5e/FlexStation® 3 not capable validated
SpectraMax® Paradigm validated contact us
SpectraMax® iD3/iD5 in review validated
perkinElmerLogo EnVision®/EnVision® Xcite validated validated
tecanLogo Infinite® M1000/M1000Pro/Safire2™ validated validated
Infinite® M200 not capable not capable
Infinite® F500 validated validated
Infinite® F200/Ultraevolution contact us contact us
Spark™ 10M validated validated

What's Included

What You Will Need

Transcreener AMP2/GMP2 Phosphodiesterase Assay Kits

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

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 play important roles in a variety of biochemical processes. PDEs are currently classified into 11 different families. Each is classed based on a number of different factors including the distribution within tissue, and substrate specificity. Some hydrolyze cAMP, while others cGMP. A variety can degrade both cAMP and cGMP. 

PDEs 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, 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

The Transcreener phosphodiesterase assay enables researchers to scour vast compound libraries for potent, yet selective inhibitors for unique PDEs. Follow-up SAR can also be completed to improve upon initial hits and provide a greater probability for success. 

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