Biology at Work
Transcreener® ADP TR-FRET Green Assay
| Product | Cat# | Size | Price |
|---|---|---|---|
| Transcreener ADP TR-FRET Green Assay | 3005-1K | 1000 assays | $585 |
| This is an evaluation kit. Sufficient reagents are provided to complete up to 1000 assays. The exact number of assays is dependent on your enzyme reaction conditions.. | |||
| Bulk Ordering | |||
| For orders of 10,000 wells or more, please contact us for a price quote. info@bellbrooklabs.com | |||
Note: BellBrook Labs has developed an improved TR-FRET Assay. For product details, please go to the Transcreener ADP2 TR-FRET Red Assay page. It incorporates our more sensitive ADP2 Antibody, and uses a far red tracer to minimize interference from fluorescent compounds and light scattering.
- Transcreener ADP TR-FRET Green Assay Technical Manual
- Transcreener ADP TR-FRET Green Assay Posters and Presentations
- Transcreener Technical References
- No Hazard Statement
Benefits
- UNIVERSAL: Homogenous ADP detection
- FLEXIBLE: Any kinase. Any substrate.
- PREDICTIVE: Rapid pharmacologically relevant data.
- SENSITIVE: Z' > 0.5 at low ATP conversion.
- SMART: A single set of reagents reduces time and expense.
The Transcreener ADP TR-FRET Green Assay is a competitive, time-resolved fluorescence resonance energy transfer (TR-FRET) immunoassay for detection of ADP. The Transcreener ADP TR-FRET Green Assay can be used for any enzyme class that produces ADP, including kinases, ATPases, DNA helicases, and ADP-glycosyltransferases. Kinases catalyze the transfer of a phosphate group from ATP to a protein, peptide, lipid, or small molecule substrate generating ADP, the invariant product. For ATPases, a phosphate group from ATP is transferred to a water molecule. The Transcreener ADP TR-FRET Green Assay was designed as a simple two-step, endpoint assay (a 10 µL enzyme reaction followed by addition of 10 µL ADP Stop and Detect Mixture, Fig. 1). The assay provides an excellent signal at low ATP conversion resulting in overall Z’> 0.6. The Transcreener ADP TR-FRET Green Assay can accommodate from 1 µM to 100 µM ATP.
The Transcreener ADP TR-FRET Green Assay can be used across an entire protein kinase family
The Transcreener ADP TR-FRET Green Assay has been developed to measure ADP and is comprised of BellBrook Labs’ monoclonal ADP Antibody conjugated to terbium (ADP Antibody-Tb) and an ADP-fluorescein (FAM) Tracer (ADP-FAM Tracer). TR-FRET is observed when the terbium chelate (covalently bound to the ADP Antibody) is in close proximity to the ADP-FAM tracer. ADP generated during the enzyme reaction displaces the ADP-FAM Tracer from the ADP Antibody-Tb and TR-FRET decreases. The Transcreener ADP TR-FRET Green Assay can be used for any enzyme class that produces ADP, including kinases, ATPases, DNA helicases, and ADP-glycosyltransferases. In addition, the generic detection of ADP is compatible within any enzyme family. For example the Transcreener™ platform detects activity of serine/threonine and tyrosine protein kinases using protein or peptide acceptor substrates, lipid kinases using lipid acceptor substrates, and metabolic kinases using carbohydrate substrates.
Step 1 Enzyme Reaction
Add 10 µL enzyme reaction to assay plate.
Step 2 ADP Detection
Add 10 µL Transcreener ADP Detection Mix, incubate 1 hour and read TR-FRET.
10 µM ADP/ATP Standard Curve Data
The 10 µM ADP/ATP Standard Curve mimics a typical enzyme reaction (as ADP is produced, ATP is depleted) by keeping the adenosine concentration constant. Graphing on the log scale eliminates the point that corresponds to zero. To include all twelve points in the curve, the 0 µM ADP/10 µM ATP was graphed at the 0.01 µM ADP/9.99 µM ATP position.
For more information on the Transcreener ADP TR-FRET Green Assay contact BellBrook Labs:
866.313.7881 in North America
info@bellbrook labs.com
Note: prior to 4/1/08 The Transcreener ADP TR_FRET Assay was known as the Transcreener KINASE TR-FRET Assay.
