The Transcreener ADP2 Assay uses a highly selective antibody and far-red fluorescent tracer to directly detect ADP produced by any ADP-producing enzyme (i.e. kinases, ATPases, helicases, carboxylases, nucleotidases, and chaperonins). The direct detection method offers a unique approach that simplifies the protocol (no coupling enzyme needed) and reduces compound interference. The assay can be used to:
- Measure activity of any ADP-producing enzyme
- Screen compound libraries for modulators
- Quantify inhibitor potency
- Inhibitor selectivity profiling
- Measure drug-target residence time
Image Credit: BellBrook Labs
Targets Validated with the Transcreener ADP2 Assay
BellBrook Labs develops complete assay solutions for tougher enzyme targets that are difficult to source enzymes and/or develop assays for (so you can focus on optimizing your lead molecules). Currently, they have assay solutions for the ADP-producing enzyme targets listed in the table below. Click on a target to learn more.
Source: BellBrook Labs
Features
- Direct detection of unlabeled ADP
- Easy to use and HTS-ready
- Highly sensitive assay reduces enzyme usage
- Generic nature makes it compatible with any ADP-producing enzyme
- Robust Assay Z' > 0.7 under initial velocity conditions
- Far-red fluorescent readouts minimize compound interference
- A safe, non-radioactive method
- Use with ATP concentrations from 0.1 to 1000 µM
- Available in FP, FI, or TR-FRET readouts
- Can be used in continuous or endpoint mode
Example Data using MAPK1 Kinase
Robust Assay Ready to Scale for HTS
Z' = 0.90. Robust data is neccessary to confirm the assay is well-suited for screening large compound libraries in a high-throughput format.
Image Credit: BellBrook Labs
Detection of ADP Under Initial Velocity Conditions
MAPK1 enzyme titration using Transcreener ADP2 Assay with FP readout. The enzyme buffer included 20 mM Tris pH 7.5, 10 mM MgCl2, 1 mM DTT, and 0.01% Tween-20. The detection mix included 1X Stop and Detect Buffer B, 6.4 μg/mL ADP2 Antibody, and 4 nM ADP2 Tracer.
Image Credit: BellBrook Labs
MAPK1 linear response. The assay shows linearity when raw data is converted to ADP using a standard curve.
Image Credit: BellBrook Labs
Screen Compound Libraries & Perform Dose Responses
MAPK1 dose response with compound staurosporine using Transcreener ADP2 FP Assay. IC50 (Staurosporine)= 0.77 μM.
Image Credit: BellBrook Labs
Lead Discovery Services
BellBrook Labs also offers lead discovery services for scientists who want to advance their program but don't want to bring a biochemical assay in-house. Their services feature customizable assay conditions, quick turnaround times, and direct collaboration with a BellBrook scientist. Lead discovery services can include:
- Inhibitor Screening
- Inhibitor Potency Profiling
- Inhibitor Selectivity Profiling
- Residence Time Measurements
- Mechanism of Action Studies
- Triage Non-Stoichiometric Inhibitors
- Compound-Target Binding with Thermal Shift Assays