Learn More About Transcreener Method Used for This TBK1 Assay

Discover TBK1 Inhibitors with the Transcreener ADP² Assay

The Transcreener ADP² Kinase Assay directly measures the ADP produced by Tank Binding Kinase 1 (also known as TBK1). By measuring the production of ADP researchers can effectively determine the activity of the TBK1 enzyme. The assay provides a powerful tool to screen entire compound libraries for TBK1 modulators to help find new therapeutics for autoimmune diseases and cancer.

The Role of TBK1 As Therapeutic Target

More than 30 known proteins are known to interact with TBK1 on various levels. One of the most studied features is the innate immune response to foreign infectious material. Toll-like receptors activate the enzyme leading to further signaling. As a regulator of the immune response pathway to viral and bacterial material, TBK1 interacts with TANK and TRAF3. TBK1 then phosphorylates interferon regulator factors (IRFs), which activate inflammatory immune response with type I IFNs. Its role in inflammatory modulation makes TBK1 a critical target for autoimmune and neurodegenerative diseases. TBK1 also regulates aspects of cell proliferation and death, making it a key focus area in cancer research.

Direct Detection of ADP to Measure TBK1 Enzymatic Activity

ADP² FP ASSAY - LEARN MORE

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

Transcreener ADP² FP Assay Tech Manual (PDF)

ADP² FI ASSAY - LEARN MORE

Positive FI signal. Compatible with simple fluorescence plate readers. Faster read time than FP or TR-FRET.

Transcreener ADP² FI Assay Tech Manual (PDF)

ADP² TR-FRET Assay - LEARN MORE

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

Transcreener ADP² TR-FRET Assay Tech Manual (PDF)

Applications

Measure Enzymatic Activity of TBK1
Screen Compound Libraries for TBK1 Inhibitors
Quantify Inhibitor Potency
Inhibitor Selectivity Profiling

Features

Direct detection of unlabeled ADP
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, FI, or TR-FRET readouts

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

The Transcreener ADP² TBK1 Assay is in a simple, HTS-ready, mix-and-read format. Run your enzyme reaction, add Transcreener reagents, and read your plates. The assay is compatible with 96, 384, and 1536-well formats.

Detection of ADP Under TBK1 Initial Velocity

The assay demonstrates linearity when raw data is converted to ADP using a standard curve. Many TBK1 interactions are currently known including phosphorylation of IRF3 and IRF7. Here we use both 10 µM ATP with 25 µM CK1tide peptide substrates. Linearity is shown here under initial velocity conditions when raw data is converted to ADP formed. Non-productive hydrolysis of ATP occurs in many kinase reactions due to many factors including autophosphorylation and water exploiting the ATP binding pocket of the enzyme.

Human TBK1 Enzyme Titration

Linear Response

Screen for Inhibitors & Perform SAR

Transcreener Assays are 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.

Dose-Response - BX-795

IC₅₀ = 1.71

Dose-Response Staurosporine

IC₅₀ = 1.14

Robust Assay Yields Quality Data

Z’ measurements using optimized TBK1 reaction conditions indicate a robust assay. Robust data like this is vital for sizeable high throughput screens that are difficult to complete due to massive sample quantity. Z' shown here = 0.74.

Three Fluorescent Readouts Available

Choose the readout that is the best fit for your lab based on preference and plate reader compatibility.

Fluorescent Intensity

Assay conditions for the TBK1 titration shown include 10 µM ATP, 10 µM CK1tide, and 10 µg/mL ADP² antibody.

TR-FRET

Assay conditions for the TBK1 titration shown include 10 µM ATP, 10 µM CK1tide, 27 nM tracer, and 8 nM ADP² Tb Antibody.

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

What's Included

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Component	Notes
ADP² Antibody	Antibody specific to the nucleotide ADP.
ADP Tracer	When displaced, changes in fluorescence can be read in an FP, FI, or TR-FRET depending on the assay format.
Stop & Detect Buffer B, 10X	200 mM HEPES (pH 7.5), 400 mM EDTA, and 0.2% Brij-35. Stops enzyme reactions requiring Mg²⁺.
ATP and ADP	ATP and ADP are used to create the ATP/ADP standard curve. ATP is used as a phosphate donor to start the reaction.

What You Will Need

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Component	Notes
TBK1 Enzyme	We have successfully used recombinant full-length human TBK1 from SignalChem Cat. #T02-10G.
Enzyme Buffer	25 mM Tris (pH 7.5), 5 mM MgCl_2,, 0.01% Brij-30. Enzyme buffer components may be optimized as required.
Substrate	Use the substrate to be phosphorylated for your experiment. Here we used CK1tide at 25 µM (Upstate Cat. #12-529)
Assay Plates	Assay Plates Can Be Purchased Directly From BellBrook Labs FP - An entirely black plate with a non-binding surface is required. We suggest Corning 384-well plates Cat. #4514 FI - An entirely black plate with a non-binding surface is required. We suggest Corning 384-well plates Cat. #4514 TR-FRET - An entirely white plate with a non-binding surface is required. We suggest Corning 384-well plates Cat. #4513

Transcreener ADP² TBK1 Assay Kits

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

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Cat. No.	Product Description	Price	Add to Cart
3013-A	Transcreener ADP2 FI Assay (200 Assay, 96 Well)	$305.00	Add to Cart
3013-1K	Transcreener ADP2 FI Assay (1,000 Assay, 384 Well)	$639.00	Add to Cart
3013-10K	Transcreener ADP2 FI Assay (10,000 Assay, 384 Well)	$3,689.00	Add to Cart
3013-100K	Transcreener ADP2 FI Assay (10 X 10,000 Assay, 384 Well)	$18,445.00	Add to Cart
3010-1K	Transcreener ADP2 FP Assay (1,000 Assay, 384 Well)	$639.00	Add to Cart
3010-10K	Transcreener ADP2 FP Assay (10,000 Assay, 384 Well)	$3,689.00	Add to Cart
3010-100K	Transcreener ADP2 FP Assay (10 X 10,000 Assay, 384 Well)	$18,445.00	Add to Cart
3011-1K	Transcreener ADP2 TR-FRET Red Assay (1,000 Assay, 384 Well)	$699.00	Add to Cart
3011-10K	Transcreener ADP2 TR-FRET Red Assay (10,000 Assay, 384 Well)	$3,989.00	Add to Cart
3011-100K	Transcreener ADP2 TR-FRET Red Assay (10 X 10,000 Assay, 384 Well)	$19,945.00	Add to Cart

Assay Plates for FI & FP

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Cat. No.	Product Description	Price
2233	Corning 96-Well Black Assay Plates (#3686), 3-Pack (200+ Assays)	$40.00	Add to Cart
2229	Corning 384-Well Black Assay Plates (#4514), 3-Pack (1,000+ Assays)	$45.00	Add to Cart
2230	Corning 384-Well Black Assay Plates (#4514), 30-Pack (10,000+ Assays)	$400.00	Add to Cart

Assay Plates for TR-FRET

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Cat. No.	Product Description	Price
2234	Corning 96-Well White Assay Plates (#3642), 3-Pack (200+ Assays)	$40.00	Add to Cart
2231	Corning 384-Well White Assay Plates (#4513), 3-Pack (1,000+ Assays)	$50.00	Add to Cart
2232	Corning 384-Well White Assay Plates (#4513), 30-Pack (10,000+ Assays)	$450.00	Add to Cart

TBK1 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 TBK1 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.

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.

In addition to our ADP² Assay, Bellbrook Labs also offers the following products to aid in the drug discovery process:

Transcreener® ADP² TBK1 Assay