Transcreener® AMP2/GMP2 FP Assay
|Application Set||Compound Screening|
1000 assays/384wells (BBL-3015-1K)
10000 assays/384wells (BBL-3015-10K)
Note: The exact number of assays depends on enzyme reaction conditions.
|Sensitivity||Excellent data quality (Z’ ≥0.7) and signal (≥100 mP polarization shift) at low substrate conversion using normal reaction conditions.|
|Range||Accommodates ATP/cAMP/cGMP concentrations ranging from1 µM to 1,000 µM.|
|Detection Type||Fluorescent polarization|
AMP2/GMP2 Antibody, AMP2/GMP2 Alexa Fluor® 633 Tracer, Tris Solution, AMP, GMP
Note: The plates are not included, please see manual for material not provided but necessary for assay.
|Other Product Data||
Click here for Complete Information from the Original Manufacturer
Easy-to-Use (mix-and-read method).Simple homogenous biochemical assay. Direct unlabeled AMP or GMP from a PDE reaction eliminates laborious coupling steps. Non-radioactive assay technique. HTS Compatible with 96, 384, and 1536-well plates. One assay, hundreds of targets!
|Declaration||Manufactured by BellBrook Labs.|
|Shipping and Handling|
|Short Term Storage||-20°C|
|Long Term Storage||-20°C|
|Handling Advice||Avoid freeze/thaw cycles.|
|Product Specification Sheet|
The Transcreener® HTS Assay platform overcomes the need for time-consuming, one-off assay development for individual members within a group transfer enzyme family by utilizing a single set of assay reagents that detect an invariant product. The generic nature of the Transcreener® HTS Assay platform eliminates delays involved in assay development for new HTS targets, and greatly simplifies compound and inhibitor profiling across multiple target families. Transcreener assays have been validated with major HTS instruments to determine the optimal filters and settings for maximal assay performance. Application notes with all important details are available.
The Transcreener® AMP2/GMP2 assay is available in two fluorescent readouts, Fluorescence polarization (FP) and Time-resolved Forster-resonance-energy-transfer (TR-FRET), which provide a safe HTS-compatible alternative to cumbersome radioassay methods and are more sensitive and less subject to interference than other detection methods. Both readouts use far-red tracers to minimize compound interference and lower false-positive rates.
The Transcreener® AMP2/GMP2 FP Assay (Prod. No. BBL-3015) is a far-red, competitive fluorescence polarization (FP) assay. The assay can be used with any enzyme that produces AMP or GMP, regardless of the substrate. Examples of enzymes include ubiquitin, small ubiquitin-related modifiers (SUMO), nucleic acid and protein ligases, phosphodiesterases (PDEs), and synthetases. The Transcreener® AMP2/GMP2 FP Assay is a simple one step homogenous detection assay, and is extremely flexible with regard to ATP/cAMP/cGMP concentrations (1 to 1,000 μM). The assay provides excellent quality with a Z’ = 0.7 and signal (≥100 mP polarization shift) at low substrate conversion (typically 10%) using normal reaction conditions.
- Development and Validation of a Transcreener Assay for Detection of AMP- and GMP-Producing Enzymes: M. Staeben, et al.; Assay Drug Dev. Technol. 8, 344 (2010)
- De-AMPylation of the small GTPase Rab1 by the pathogen Legionella pneumophila: M.R. Neunuebel, et al.; Science 333, 453 (2011)
- Development and validation of a generic fluorescent methyltransferase activity assay based on the transcreener AMP/GMP assay: T.A. Klink, et al.; J. Biomol. Screen. 17, 59 (2012)
- Selective and specific inhibition of the plasmodium falciparum lysyl-tRNA synthetase by the fungal secondary metabolite cladosporin: D. Hoepfner, et al.; Cell Host Microbe 11, 654 (2012)
- Identification of potent inhibitors of the Trypanosoma brucei methionyl-tRNA synthetase via high-throughput orthogonal screening: L. Pedró-Rosa, et al.; J. Biomol. Screen. 20, 122 (2015)
- Fluorescence polarization immunoassays for monitoring nucleoside triphosphate diphosphohydrolase (NTPDase) activity: A. Fiene, et al.; Analyst 140, 140 (2015)
- Polyoxometalates--potent and selective ecto-nucleotidase inhibitors: S.Y. Lee, et al., Biochem. Pharmacol. 93, 171 (2015)