Transcreener® OAADPr SIRT Assay Kits

Download Tech Manuals

FP Manual TR-FRET Manual

Single-Step SIRT Assay for Screening and Lead Discovery

Simple - mix-and-read HTS-ready format
Save time and money with homogenous validated assays
Detection of unlabeled OAADPr - Instead of fluorophore-tagged peptide substrates.
Robust Assay Z’ > 0.7 under initial velocity conditions
Flexible - Use for inhibitor screening, dose-response assays, kinetic, and mechanistic analysis

Is your plate reader compatible? Find out now!

Custom Sizes and Bulk Pricing Available Contact Us for an HTS-scale Quote!

Readout

Assay

Clear

SKU: N/A Category: Assays

Overview – Transcreener OAADPr SIRT Assay

Name: Transcreener® OAADPr SIRT Assay Kits
SKU: 27051
Availability: InStock

The Transcreener OAADPr SIRT Assay is a biochemical HTS assay for measuring the production of 2’-O-acetyl-ADP-ribose (OAADPr) by sirtuins (SIRT), NAD-dependent deacetylases.

The kit comes complete with the detection reagents required to measure activity. SIRT enzymes are not included in the inhibitor screening assay kit. Recombinant SIRT1 and SIRT2 enzyme and Enzolution SIRT1, SIRT2, and SIRT3 Assay Systems are available for separate purchase through BellBrook labs.

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 OAADPr Assay Work?

The Transcreener OAADPr SIRT Assay is a biochemical HTS assay for measuring the production of 2’-O-acetyl-ADP-ribose (OAADPr) by sirtuin (SIRT) reactions.

The assay relies on a Coupling Enzyme (CE) to convert OAADPr into AMP, which is then detected using a far-red, competitive fluorescence assay with an FP or TR-FRET readout. As an example, the Transcreener OAADPr SIRT Assay can be used to detect the activity of any human SIRT using acetylated peptides as substrates.

The Transcreener OAADPr SIRT Assay is designed specifically for high throughput screening (HTS), with a single-addition, mix-and-read format.

Transcreener OAADPr SIRT Assay Schematic v5.23.25

It is easy to integrate into automated HTS workflows, with outstanding reagent stability (deck stability > 16 hours, signal stability > 24 hours), robust detection of OAADPr, and compatibility with commonly used multimode plate readers. Data quality is excellent (Z’ ≥ 0.7), and the assay uses a far-red tracer to minimize interference from fluorescent compounds and light scattering.

Overcoming SIRT Assay Limitations: Fast, Homogeneous Detection of OAADPr

In this video, we introduce a new universal Sirtuin activity assay that measures OAADPr in real time by enzymatically converting it to AMP, which is detected using the Transcreener AMP Assay with either an FP or TR-FRET readout. This innovative method eliminates the complexity of substrate-specific readouts and enables reliable, quantitative measurement of SIRT enzymatic activity for any substrate and any Sirtuin.

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

Combine with the Enzolution SIRT Assay Systems to Find Novel Inhibitors in Hours Not Months

What’s Included

FP
	1K		10K
Component	Part #	Qty	Part #	Qty
AMP²/GMP² AlexaFluor® 633 Tracer, 800 nM	2097	100 μL	2101	1 mL
AMP²/GMP² Antibody, 1.26 mg/mL	2114	158 μL	2344	1.6 mL
OAADPr Coupling Enzyme, 200X	2334	50 μL	2335	500 μL
NAD+, 5 mM	2330	300 μL	2331	3 mL
AMP, 5 mM	2099	200 μL	2099	200 μL
Stop & Detect Buffer B, 10X	2027	1 mL	2032	10 mL
Enzyme Assay Buffer G, 10X	2338	2 mL	2339	20 mL
DTT, 1M	2296	100 µL	2297	1 mL
MnCl₂, 1M	2303	30 µL	2304	300 µL

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
OAADPr Coupling Enzyme, 200X	2334	50 µL	2335	500 µL
NAD+, 5 mM	2330	300 μL	2331	3 mL
AMP, 5 mM	2099	200 μL	2099	200 μL
Stop & Detect Buffer B, 10X	2027	1 mL	2032	10 mL
Enzyme Assay Buffer G, 10X	2338	2 mL	2339	20 mL
DTT, 1M	2296	100 µL	2297	1 mL
MnCl₂, 1 M	2303	30 µL	2304	300 µL

What You Will Need

Component	Notes
Ultrapure Nuclease Free Water	Some deionized water systems are contaminated with enzymes that can degrade both nucleotide substrates and products, reducing assay performance. Use nuclease free water such as: Invitrogen Part # AM9930
Enzyme	You can purchase active *SIRT1 enzyme, SIRT2 enzyme, and SIRT3 enzyme* directly through BellBrook Labs.
Acetylated Peptide	The assay has been validated using H3K9Ac (1-21), an acetylated peptide derived from the N-terminus of Histone H3. H3K9Ac (1-21) is supplied in SIRT1, SIRT2, and SIRT3 Enzolution Assay kits (Part # 3048, 3049, and 3052).
Plate Reader	A multi-detection plate reader configured to measure the output of the tracer is required.
Liquid Handling Devices	Use liquid handling devices that can accurately dispense sub-microliter volumes into 384-well plates.
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).

Applications

Measure Enzymatic Activity of SIRTs
Screen Compound Libraries for SIRT inhibitors
Quantify Inhibitor Potency – Generate Accurate IC₅₀s
Kinetic and Mechanistic Analyses

Features

Detection of unlabeled OAADPr produced by SIRTs
Easy to use, homogenous, one-step format
Robust Assay Z’ > 0.7 under initial velocity conditions
Far-red fluorescent readouts minimize compound interference
A safe, non-radioactive method
Available in FP or TR-FRET readouts
Less than 0.1% interference in tested libraries

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

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

Transcreener OAADPr SIRT Assay FP Outline

Detection of OAADPr to Measure SIRT1 Enzymatic Activity

SIRT1 enzyme titration using the Enzolution SIRT1 Assay System. The assay demonstrates linearity when raw data is converted to AMP using a standard curve.

Human SIRT1 Titration

Linear Response

Robust HTS Ready Assay to Measure SIRT1 Enzymatic Activity

Z’ of 0.80 or greater confirms the robustness of the SIRT1 assay. A robust system like this is necessary to ensure the precision and efficiency required to scale up assays for high-throughput screening. No need to scale up on your own!

FP Readout

TR-FRET Readout

Screen for Inhibitors & Perform SAR

The Transcreener OAADPr SIRT Assay is designed for screening compound libraries in a high throughput format. Follow up SAR can also be performed using the assay to determine inhibitor potency with ease.

Tocris 2.0 Library Pilot Screen with SIRT1

Accurate Dose Response Results with SIRT Inhibitors

Example dose-response curves with SIRT1 and SIRT2 inhibitors. Data was generated using the Transcreener OAADPr SIRT FP Assay, with the Enzolution SIRT1 or SIRT2 Assay Systems. The following conditions were used. Assay buffer G with 0.5 mM MnCl₂, 1 mM DTT, 0.01% Triton, 50 µM NAD+, 100 μM H3K9Ac Peptide, with 1X OAADPr coupling enzyme mix. Either 3 nM SIRT1 or 12 nM SIRT2 was used. The enzyme was incubated with the test compound for 30 minutes before the addition of CE/substrate mix and allowed to incubate for 60 minutes. The 1X AMP detection mix was added and incubated for 120 minutes, and the plates were read via a plate reader. EX-527 is available for purchase through BellBrook Labs.

SIRT1 with EX-527

IC₅₀ = 206 nM

SIRT2 with A-419259 3HCl

IC₅₀ = 0.71 µM

SIRT3 with 3-TYP

IC₅₀ = 27.17 µM

Recombinant SIRT1 Enzyme

Active, full-length human SIRT1 (AA 1-747) protein expressed and purified from Baculovirus infected insect cells with a C-Terminal His tag. The enzyme has been thoroughly validated with the Transcreener OAADPr Assay Kit. For specific activity, please refer to the Certificate of Analysis for each individual enzyme lot. Purity is >85% in a composition of 50 mM Tris, 500 mM NaCl, and 5% Glycerol (pH 8.0). Currently used with the Transcreener assay at a concentration of 3 nM in a 10 μL reaction.

Go to Human SIRT1 Enzyme Page

Recombinant SIRT2 Enzyme

Active, full-length human SIRT2 (AA 1-389) protein expressed and purified from Baculovirus infected insect cells with a C-Terminal His tag. The enzyme has been thoroughly validated with the Transcreener OAADPr Assay Kit. For specific activity, please refer to the Certificate of Analysis for each individual enzyme lot. Purity is >90% in a composition of 50 mM Tris, 500 mM NaCl, and 5% Glycerol (pH 8.0). Currently used with the Transcreener assay at a concentration of 12 nM in a 10 μL reaction.

Go to Human SIRT2 Enzyme Page

Recombinant SIRT3 Enzyme

Active, human SIRT3 (AA 101-399) protein expressed and purified from E.Coli with an N-Terminal His tag. The enzyme has been thoroughly validated with the Transcreener OAADPr Assay Kit. For specific activity, please refer to the Certificate of Analysis for each individual enzyme lot. Purity is 95% in a composition of 50 mM HEPES, 200 mM NaCl, 1 mM TCEP, 1 mM EDTA, 10% Glycerol, pH 7.5. Currently used with the Transcreener assay at a concentration of 24 nM in a 10 μL reaction.

Go to Human SIRT3 Enzyme Page

Sirtuin Assay Services

Interested in moving your Sirtuin program forward, but don’t want the hassle of bringing an assay in-house? BellBrook’s expert scientists have you covered. Our team of biochemical and enzymology expert scientists will work diligently to progress your lead candidates through the early-stages of drug discovery. The best part – we are a small custom shop. We can customize assay conditions to suit your project needs, produce results faster, and you will directly collaborate with one of our expert scientists.

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.
Mechanism of Action Studies – Kinetic analysis to define the mode of inhibition.
Inhibitor Screening – To identify or confirm activity with the target.
Triaging Non-stoichiometric Inhibitors – Run assays under different conditions, such as varying detergent and enzyme concentrations.
Evaluate Compound-Target Binding – Run thermal shift assays to confirm compound-target engagement via shifts in melting temperature.

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.

The Role of Sirtuins As Therapeutic Targets

SIRTs, or Sirtuins, are a family of enzymes crucial to various cellular processes, including aging and metabolism. As NAD+-dependent deacetylases, they require NAD+ to remove acetyl groups from proteins, influencing cellular functions.

Particularly SIRT1 has gained attention as a therapeutic target due to its role in conditions associated with aging, metabolic disorders, cardiovascular diseases, and neurodegenerative diseases. SIRT1 helps promote DNA repair, regulate metabolic pathways, and enhance stress resistance, contributing to cellular longevity.

Modulating sirtuin activity, especially SIRT1, may help extend lifespan and delay age-related diseases like Alzheimer’s and Parkinson’s. Ongoing research aims to develop small molecules or biologics to effectively target sirtuins, showcasing promise for various therapeutic applications.

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

Previous Next

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

info@bellbrooklabs.com

Also required (not included):

Enzolution® SIRT2 Assay System

Enzolution® SIRT1 Assay System

Enzolution® SIRT3 Assay System