Transcreener® ADP² TBK1 Assay

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

ADP FP Kinase Assay

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

ADP FI Kinase Assay

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

ADP TR-FRET ATPase Assay

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

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, FI & TR-FRET Readouts Validated on Major Multimode Readers

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

What's Included

What You Will Need

Transcreener ADP2 TBK1 Assay Kits

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

Assay Plates for FI & FP

BellBrook Labs
5500 Nobel Drive, Suite 230
Madison, Wisconsin 53711 USA
(608) 443-2400

info@bellbrooklabs.com

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