Biology at Work
Transcreener® ADP Assay- far red FP
Note: prior to 4/1/08 The Transcreener ADP Assay was known as the Transcreener KINASE Assay.
| Product | Cat# | Size | Price |
|---|---|---|---|
| Transcreener ADP Assay | 3004-1K | 1000 assays | $450 |
| Bulk Ordering | |||
| For orders of 10,000 wells or more, please contact us for a price quote. info@bellbrooklabs.com | |||
*The Transcreener ADP Assay is a highly flexible format and can detect enzyme activity over a wide range of starting ATP concentrations (1 µM to 1,000 µM) in diverse reaction buffers. Sufficient reagents are provided in 3004-1K to complete the protocol when using ATP within a range of 1 µM to 100 µM ATP. If you are using >100 µM ATP please call BellBrook Labs for custom packaging.
If you are measuring fluorescence polarization with the Perkin Elmer EnVision™ multilabel plate reader, please tell us when placing your order as this instrument requires modified assay optimization conditions. Further, if you are using 1536 well plates, modified assay optimization conditions may enhance performance. Please contact BellBrook Labs for additional details.
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- Transcreener ADP Assay Technical Manual
- Transcreener ADP Assay Posters and Presentations
- Frequently Asked Questions
- A Guide to Optimizing Assay Window and Sensitivity
- Transcreener ADP Assay Case Study: PKA Inhibitor Screen
- No Hazard Statement
Benefits
- UNIVERSAL: Homogenous ADP detection
- FLEXIBLE: Any ADP-producing enzyme and substrate
- RELEVANT: Rapid, pharmacologically relevant data
- SENSITIVE: Z' > 0.7 at low ATP conversion.
- SMART: One set of reagents reduces time and expense.
The Transcreener ADP Assay can be used across an entire group transfer enzyme family
A far-red competitive fluorescence polarization assay that detects ADP produced during the transfer of phosphate (from ATP) to acceptor substrate. Because the technology relies on detection of the invariant reaction product, a single set of reagents can be used for any ADP producing enzyme and any peptide or native substrate. The Transcreener ADP Assay is not limited to use with traditional kinase enzymes, but can be used to assay all ADP-producing enzymes, such as protein, lipid and carbohydrate kinases, ATPases, DNA helicases and carboxyltransferases.
Flexibility for a wide range of ATP concentrations
The Transcreener ADP Assay can be used over a wide range of ATP concentrations. In an attempt to mimic ADP generated during a kinase reaction, standard curves optimized for 1 µM and 200 µM ATP are prepared by keeping the adenosine concentration constant. The IC50 values for the 1 µM, 10 µM, and 200 µM ATP-ADP curves are 0.3 µM, 1.3 µM, and 20 µM ADP, respectively, showing that a low concentration of ADP is capable of producing significant shifts in mP values.
Accelerate Kinase Profiling
Profiling of known inhibitors with PKA, cdk5/p35 and Abl kinases demonstrates correct pharmacology using the Transcreener ADP Assay. A panel of twelve kinase inhibitors at 10 µM (1% DMSO) were incubated in 10 µM ATP and 40 ng/mL Abl or 240 ng/mL cdk5/p35 in the Transcreener ADP Assay. At these enzyme concentrations, approximately 30% of the ATP was converted to ADP. As expected, Staurosporine inhibited all three kinases, while Rotterlin did not inhibit kinase activity. Gleevec® and Tryphostin AG1478 specifically inhibited Abl, whereas H89 and Go-6983 wer selective towards PKA. Roscovitine only inhibited cdk5/p35 in the absence of Histone H1. The inhibitor panel also identified correct pharmacology for Akt/PKBalpha kinase at 75µM ATP, the ATP Km for the enzyme (data not shown).
For more information on the Transcreener ADP Assay contact BellBrook Labs:
866.313.7881 in North America
info@bellbrook labs.com