BellBrook Labs, a US-based company, is dedicated to accelerating drug discovery and biological research. The company’s Transcreener^® HTS enzyme assays, used by all of the major pharmaceutical companies, make it easy to screen thousands of different enzymes, including validated targets like kinases, as well as emerging targets like ATPases, GTPases, methyltransferases, cGAS, Helicases, and glycosyltransferases. Assay development and lead discovery services help customers advance targets through drug discovery programs with confidence.

BellBrook Labs Transcreener^® and AptaFluor Assay Kits are robust, repeatable, easy-to-use enzyme activity assays for drug discovery. By directly detecting the common product of an enzymatic reaction, e.g. ADP for kinases, Transcreener provides a universal method that can be used across an entire target family. Direct detection and far-red fluorescent readouts (by fluorescence polarization, fluorescence intensity or TR-FRET) limit interference, producing accurate results that accelerate your drug discovery.

AdipoGen Life Sciences provides the complete BellBrook Labs product range in Switzerland and Italy.

For a product overview please see below:

• Transcreener® & AptaFluor Assay Technology
• NEW & UNIQUE Transcreener® cGAMP cGAS Assay
• Transcreener® and Aptafluor Assays Overview
• Enzolutions™ - Validated Assay Overview
• Validated Active Enzymes
• Corning Plates for Transcreener Assays

For Product Inquiries: info@adipogen.com

Transcreener^® Assay Technology: How does it work?

Transcreener^® is a universal HTS assay method that can be used across entire families of nucleotide-dependent enzymes and relies on direct immunodetection of nucleotides. The panel of assays for ADP, GDP, AMP, GMP, cGAMP, dAMP, ADPR, 2-5A, SAH, and UDP measure activity for thousands of enzymes.

Transcreener^® assays detect highly specific nucleotides using antibodies that can differentiate between nucleotides based on a single phosphate group. Selectivities for the product nucleotides vs. the substrates (e.g., ADP vs. ATP for a kinase assay) range from 150-fold to over 1000-fold. This exquisite selectivity allows detection of nucleotide enzyme products in the presence of an excess of the substrate nucleotide (e.g, ADP detection in the presence of excess ATP for kinase assays), a requirement for measuring enzyme initial velocity.

To generate a signal, the Transcreener^® assays use a homogenous, competitive immunoassay format in which the antibodies are paired with high-affinity fluorescent tracers. Displacement of the tracer by the nucleotide being detected causes a change in its fluorescence properties. Transcreener provides a choice of three fluorescent readouts; FP, TR-FRET, and FI.

Transcreener^® assays have fewer reagents and a less complex mechanism than any other nucleotide detection assay, which generally require coupling enzymes to convert a nucleotide to a product that can generate a signal with a reporter enzyme. Each coupling and reporter enzyme 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.

Direct detection also means that the protocol is the simplest and most flexible available. They are validated on all major multimode readers with well-defined filters and settings (for Instrument Compatibility Table) and can be used with 96, 384, or 1536 well formats.

Thousands of cellular reactions use nucleotides, including most of the enzymes that catalyze posttranslational modifications of proteins such as phosphorylation and methylation. Because of their central role in signal transduction, many PTM enzymes have become drug targets. Though protein kinases get the most attention, acetyltransferases, methyltransferases, and glycosyltransferases are also clinically validated targets and many more PTM-related drugs are in the pipeline. Transcreener^® assays were developed to enable HTS efforts targeting PTM enzymes, as well as other types of nucleotide-dependent enzymes, including ATPases, GTPases, and phosphodiesterase (for Application Notes).

Comparison between the Transcreener ADP2 Assay from Bellbrook Laboratories and the ADP-Glo Assay from Promega, show multiple advantages of the Transcreener assay (for Comparison Slides).

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AptaFluor – Leveraging the Power of Aptamers for Drug Discovery

Epigenetic regulation has been shown to be a contributing factor in a variety of diseases. The discovery of methyltransferase inhibitors is currently an area of intense activity. Methyltransferase enzymes methylate a variety of substrates using S-adenosylmethionine (SAM) as the methyl donor leaving the common product S-adenosylhomocysteine (SAH).

The AptaFluor SAH Methyltransferase Assay uses a naturally occurring aptamer, or riboswitch, that selectively binds with SAH, the invariant product of methyltransferase reactions. The exquisite affinity and selectivity of the riboswitch combined with a positive TR-FRET signal enable screening and profiling of methyltransferases with unparalleled sensitivity.

By directly detecting SAH in a homogenous format, it is possible to perform screening and enzymatic studies with virtually any methyltransferase. The assay can be used as a tool to aid researchers in hit identification and characterization in a quest to develop new methyltransferase inhibitors as treatments for disease.

Key Attributes/Advantages:

• Direct Detection of SAH with a TR-FRET Readout
• Ultra-Sensitive
• Use with Physiological SAM Concentrations Lower than 100nM
• Outstanding Reagent and Signal Stability
• Excellent Z’ under a variety of assay conditions
• Simple Mix-and-Read
• Universal Assay – Use with Virtually Any Methyltransferase

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Transcreener^® and AptaFluor Assay Kits can be used for:

• 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

Important Advantages to Competitive Assays are:

• Universal – Use with virtually any kinase
• Direct detection of nucleotide enzyme products eliminates complex coupling steps that add to assay interference
• The sensitivity of Transcreener enables accurate data, generating Z’ values of greater than 0.7 at substrate conversion levels less than 10%
• Saves you time and development costs by using a single set of reagents for the entire kinase enzyme family
• Your choice of FP, FI, and TR-FRET readouts with certified performance on major multimode readers
• Overnight reagent and signal stability means you get reliable, robust screening data in large automated screens
• Safe, non-radioactive assay method
• Measure enzyme activity in real-time allowing for residence time determination in an HTS format.

For Detailed information please see the Transcreener^® HTS Assay Overview Page and download BellBrook Labs Guide to "Measuring Drug-Target Residence Times with Biochemical Assay"

For FAQs

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NEW & UNIQUE Transcreener^® cGAMP cGAS Assay

The Transcreener^® cGAMP cGAS FP Assay (Prod. No. BBL-3024) is a far-red, competitive fluorescence polarization (FP) assay. Because the antibody is highly selective for cGAMP, the assay can be used to measure activity of the cyclic GMP-AMP synthase (cGAS) enzyme which converts ATP and GTP, to cGAMP. cGAS is a recently discovered enzyme that acts as a foreign DNA sensor that induces an immune response via activation of the stimulator of interferon genes (STING) receptor. By directly measuring cGAMP with a highly selective antibody, it is possible to assay the activity of cGAS while screening large compound libraries for inhibitors. This assay can be used to i) measure enzymatic activity of cGAS, ii) screen compound libraries for cGAS inhibitors, iii) quantify inhibitor potency or iv) determine inhibitor-cGAS residence time. 

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Transcreener^® and Aptafluor Assays Overview

Transcreener® is the simplest, most robust HTS platform available for nucleotide-dependent enzymes. These are Mix-and-Read assays for Kinases, ATPases, GTPases, methyltransferases, phosphodiesterases, glycosyltransferases, DDR enzymes and immune sensors with first-in-class sensitivity and performance.

• The only HTS method for direct detection of nucleotides produced by enzymes
• Detects thousands of enzymes: kinases, ATPases. GTPases, phosphodiesterases, glycosyltransferases, exonucleases, etc.
• Continuous OR end point assays
• True single-step mix-and-read format

AptaFluor is the simplest, most sensitive methyltransferase assay available.

• Mix-and-read SAH detection
• Detect any methyltransferase using SAM at Km concentration
• Use with virtually any methyltransferase enzyme

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Enzolutions^™ - Validated Assay Overview

Enzolution™ is a complete assay solution. Here is how it works. You take a Transcreener Assay and an Enzolution™ Assay System (sold separately), and together you have everything you need perform inhibitor screening and dose-response measurements for a given target. This includes assay plates, substrates, enzymes, and assay buffers. Enzymes are also available individually!

• Validated enzymes, substrates, buffers, and protocols, ready for inhibitor discovery
• Designed to be used with the Transcreener and AptaFluor platforms
• Enzolution™ is the one-stop-shop for your biochemical assay needs

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Validated Active Enzymes

Highly active human enzymes validated for HTS in Bellbrook Laboratories proprietary assays: cGAS, TREX1, PolQ, WRN, PARG, CD38, DDX helicases, MDA5, RIG-I, and many more.

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Corning Plates for Transcreener Assays - Choose the Right Plate for your Assay

Transcreener® HTS Assays rely on direct immunodetection of nucleotides with a far-red readout in a simple mix-and-read format. The assays are available in three different detection modes, fluorescence polarization (FP), fluorescence intensity (FI), and time-resolved Forster resonance energy transfer (TR-FRET). A good microplate reader and the right choice of plates to differentiate specific signals versus background signals or “noise” is crucial to the success of these assays.

To achieve the best results be sure to:

- Use polystyrene, non-binding plates
- Make sure the plate has the correct working volume
- Use black plates for FP & FI
- Use white plates for TR-FRET

 

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