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Measuring Kinase Inhibitor Residence Times with the Transcreener® ADP2 FP Assay

BellBrook Labs

This Application Note provides a protocol, experimental examples, and data analysis guidelines for using the Transcreener® ADP2 FP Assay to determine dissociation rates and residence times for kinase inhibitors using simple HTS-compatible methods.

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Case Studies

Measuring Phosphodiesterase Inhibitor Residence Times with the Transcreener® AMP2/GMP2 FP Assay

BellBrook Labs

This Case Study uses a jump-dilution method with the Transcreener® AMP2/GMP2 FP Assay to determine the residence times for PDE inhibitors.

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Discovery of GALNT3 Inhibitors Using a Universal Glycosyltransferase Assay and Orthogonal Pooled Screening

BellBrook Labs

This Case Study shows how to develop an HTS workflow for glycosyltransferases (GTs) with an optimized Transcreener® UDP2 Assay (FP and TR-FRET formats), use the assay to measure GT activity and determine Km values, run a pilot screen for specific enzyme inhibitors on 8,000 compounds using Orthogonal Pooled Screening (OPS™) technology, and confirm, validate, and characterize hits.

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Exploring the Sumoylation Cascade using the Transcreener® AMP2/GMP2 Assay

BellBrook Labs

This Case Study helps you learn how to detect AMP formed by E1 alone or the complete SUMO ligase cascade in a homogenous, mix and read assay and determine the potency of SUMO ligase inhibitors.

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Selective Detection of LRRK2 GTPase Activity with the Transcreener® GDP Assay

BellBrook Labs

This Case Study helps you learn how BellBrook Labs developed a selective and sensitive, HTS-ready assay for the GTPase domain of LRRK2 using the Transcreener® GDP FP Assay.

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Customer Success Story:
Development of a High-Throughput Screening Assay to Identify Inhibitors of the de novo Purine Biosynthetic Pathway

David Whalley et al.

This Case Study helps you learn more about David Whalley and his success using the BellBrook Labs Transcreener® ADP assay. David leveraged the Transcreener® ADP assay to identify inhibitors of PAICS, a key enzyme in the de novo purine biosynthesis pathway that has been suggested as an attractive drug target for cancer.

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Tutorials

Orthogonal Pooled Screening: HTS with 5X higher efficiency, using 5X fewer resources

BellBrook Labs

In this Tutorial, explore what orthogonal pooled screening (OPS™) is, how the OPS™ Platform works, why you should consider using the OPS™ Platform and the LCGC library, and what benefits the OPS™ Platform provides in terms of efficiency, resources, and results.

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Homogenous Transcreener® UDP2 TR-FRET Assay for High Throughput Screening with Glycosyltransferases

BellBrook Labs

In this Tutorial, learn how to use Transcreener® HTS assays for direct detection of glycosyltransferases that use different sugar-nucleotide donors, how to measure enzyme activity and inhibitor dose responses for several GTs, including human fucosyltransferase, glucosylceramide synthase, galactose N-acetyltransferases, β1-4 galactosyltransferase, sialyltransferase and C. dificile Toxin B.

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Transcreener® Assay Development Services

BellBrook Labs

In this Tutorial, learn about our Assay Development Services, including our quick enzyme activity test, options for assay optimization and inhibitor profiling, and pilot screen capabilities.

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Profiling Phosphodiesterases with the Transcreener® AMP2/GMP2 TR-FRET Assay

BellBrook Labs

In this Tutorial, learn about our Transcreener® AMP2/GMP2 TR-FRET Assay, with a highly specific antibody and a simple, mix-and-read format, lighting the way in cyclic nucleotide phosphodiesterase profiling.

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Transcreener® ADP2 Assays for universal screening and profiling of any kinase or ATPase

BellBrook Labs

In this Tutorial, learn about our Transcreener® ADP2 Assays and their single addition, mix-and-read assay format compatible with automated high throughput screening and profiling protocols, the reliable Transcreener® assay reagents and assay signals that simplify your workflows, how enzyme costs are reduced and fewer false positives are observed.

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Probing p97 ATPase with the Transcreener® ADP2 ATPase Assay

BellBrook Labs

In this case study, BellBrooks’ scientific team optimized enzyme reaction and detection conditions that produced outstanding assay windows and Z’ values for both forms of the enzyme within one week.

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Streamlining Kinase Screening and Profiling with Transcreener® ADP2 Assays

BellBrook Labs

In this Tutorial, learn about our Transcreener® ADP2 Assays, our instrument validation program to insure optimal performance on all major multimode readers, calibration line for ATP concentrations from 0.1 to 1,000 μM, use of EC40–EC80 enzyme concentration, and the ability to determine accurate inhibitor IC50 values directly from raw fluorescence data, eliminating the need for standard curves.

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Transcreener® HTS Assays for Epigenetic Enzymes

BellBrook Labs

In this Tutorial, learn how each of these four Transcreener® assays enables detection of epigenetic enzymes in a robust, HTS-ready format: ADP2 Assay for kinases, AMP2/GMP2 Assay for Ub and Sumo ligases, EPIGEN Methyltransferase Assay for HMTs, and EPIGEN Acetyltransferase Assay for HATs.

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Validation of Reaction Biology’s recombinant human methyltransferases in the Transcreener® Epigen Methylatransferase Assay

BellBrook Labs

In this Tutorial, you can learn how BellBrook’s Transcreener® EPIGEN MT Assay provides universal detection of histone and DNA methyltransferases with the sensitivity and robustness needed for industrial HTS as well as how we are determining the optimal reaction conditions to produce an outstanding assay window under initial velocity conditions for each of RBCs enzymes, including challenging targets such as NSD2, MLL4 and EzH2.

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Screening for inhibitors of RGS protein GAP activity using the RGScreen Assay

BellBrook Labs

In this Tutorial, you can learn how BellBrook overcame the technical barriers to enable screening for RGS inhibitors in a direct biochemical HTS assay format, why RGS proteins are emerging as an exciting new class of drug targets, how RGS GAP activity can be easily measured in a homogeneous fluorescent assay format, and how 7 RGS4-specific inhibitors were identified from a 960 compound pilot screen.

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Targeting the cGAS-STING Pathway Using a Homogenous, HTS Compatible Transcreener® cGAS Assay

BellBrook Labs

In this poster, you will learn how you can use the Transcreener cGAMP cGAS assay to interrogate cGAS modulators with an easy-to-use, HTS ready, mix-and-read assay.

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A High Throughput Assay for the Ectonucleotidase CD39 Based on the Transcreener® Assay Platform

BellBrook Labs

In this poster, you will learn how you can use the Transcreener AMP assay to interrogate CD39 modulators with an easy-to-use, HTS ready, mix-and-read assay.

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Optimization of Enzyme Reactions for Screening and Profiling Wild Type EZH2 with the Transcreener EPIGEN Methyltransferase Assay

BellBrook Labs

In this poster, you will learn how you can use the Transcreener EPIGEN assay to interrogate EZH2 modulators with an easy-to-use, HTS ready, mix-and-read assay.

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The Transcreener CD73 Assay: A High Throughput, Sensitive Assay to Measure Adenosine

BellBrook Labs

In this poster, you will learn how you can use the Transcreener CD73 Assay to discovery novel inhibitors for the enzyme. See how the assay was used in a pilot screen, for dose-response curves, and more!

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Targeting ENPP1 for Cancer Immunotherapy: Development of an HTS Method Using the Transcreener® AMP2/GMP2 Assay

In this poster, you will learn how you can use the Transcreener AMP2/GMP2 Assay to discover novel inhibitors for ENPP1. See how the assay was used with different substrates including ATP and cGAMP as an HTS tool for drug discovery.

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Targeting OAS Enzymes with the Transcreener® 2-5A OAS Assay

In this poster, you will learn how you can use the Transcreener 2-5A Assay to discover novel modulators for OAS enzymes See how the assay was developed, and how it was used to screen OAS1 and OAS2, and used for SAR studies.

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A Universal Mix-and-Read Transcreener UDP FI Glyocsyltransferase Assay to Study GALNT Actvity

In this poster, you will learn how you can use the Transcreener UDP Assay to discover novel modulators for GALNT enzymes. See how the assay measures GALNT2 and GALNT3 enzymatic activity with an easy-to-use, mix-and-read, HTS-read method.

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Sensitive Detection of PRMT4 Using Physiological SAM with the AptaFluor® Methyltransferase Assay

Here we compare the AptaFluor Methyltransferase Assay to two other HTS-compatible HMT assay methods based on a) immunodetection with an HTRF® readout and b) a coupled enzyme assay with a luminescent readout. We used PRMT4 to compare the assays for detection of HMT activity, as this enzyme is representative of typical low Km HMTs (Km = 140 nM) and it has attracted interest as a therapeutic target.

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AptaFluor SAH: A Homogenous, Universal Methyltransferase Assay Based on a Microbial Riboswitch

HMTs are slow enzymes, and current SAH detection methods are not sufficiently sensitive to allow detection of many HMTs using physiological concentrations of SAM. To overcome this technical gap, we leveraged a naturally occurring SAH-sensing RNA aptamer, or “riboswitch”, that binds SAH with nanomolar affinity and exquisite selectivity. Here, we used the AptaFluor SAH Methyltransferase Assay to detect purified HMTs at concentrations well below the sensitivity limit for current assays using diverse acceptor substrates ranging from peptides to intact nucleosomes.

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

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