Direct AMP/GMP Detection method with a TR-FRET readout

Homogenous detection of AMP or GMP producing enzymes (e.g. ligases, synthetases, phospodiesterases) is now possible  with a TR-FRET readout. Like the original, FP-based assay the Transcreener® AMP2/GMP2 TR-FRET assay is designed for automated HTS platforms:run your enzyme reaction, add detection reagents, and read the far red TR-FRET signal on any multimode plate reader using the same filter sets as HTRF®.

XMP enzyme products, including AMP, GMP and CMP displace a fluorescent tracer from a highly selective antibody resulting in a decrease in FRET. The time gated detection and far red tracer reduce compound interference and provide a robust ratiometric readout.

One Assay … Hundreds of targets

  • Ligases: Ub and SUMO Ligases, DNA and RNA Ligases
  • Synthetases: Acyl CoA Synthetase, Amino-Acyl tRNA Synthetases, NAD Synthetase
  • Phosphodiesterases: cAMP, cGMP
  • Sialyltransferases: (CMP detection)
  • Protein deAMPylase
  • SINGLE ADDITION: one-step, simple mix and read format.
  • ROBUST: Z’ > 0.7 for initial velocity reactions
  • LESS INTERFERENCE: Far red fluorescence minimizes compound interference.

The instrument validation program is a collaborative effort with the major suppliers of multimode readers to ensure that Transcreener assays perform optimally on your specific instrument. Scientists at BellBrook and/or the instrument vendor have tested Transcreener reagents with each instrument to determine the optimal filters and settings for maximal assay performance.  We provide you with application notes with all of the important details, so you can be confident of success using Transcreener assays on the instrument of your choice.

Supplier Instrument TR-FRET Assays
ADP2, AMP2/GMP2, GDP, UDP, AptaFluor SAH
bioteklogo TriStar²S LB 942 validated
Mithras² LB 943 validated
bioteklogo Cytation™ 5 validated
Cytation™ 3 validated
Cytation™ 1 validated
Synergy™ H1 validated
Synergy™ 2/H4/4 validated
Synergy™ HTX not capable
Synergy™ Neo 2 validated
BMGLABTECH Logo POLARstar® Omega validated
FLUOstar® Omega validated
PHERAstar® FSX validated
PHERAstar® Plus/FS validated
CLARIOstar® /Plus validated
VANTAstar validated
MDS AT logo Analyst® GT/HT validated
Gemini® XPS/EM not capable
SpectraMax® M2/M2e not capable
SpectraMax® M5/M5e/FlexStation® 3 validated
SpectraMax® Paradigm contact us
perkinElmerLogo EnVision®/EnVision® Xcite validated
tecanLogo Infinite® M1000/M1000Pro/Safire2™ validated
Infinite® M200 not capable
Infinite® F500 validated
Infinite® F200/Ultraevolution contact us
Spark™ 10M validated

Validation Criteria

  • 384-Well Format
  • Z’-Factor ≥ 0.7 at 10% conversion of 10 μM ATP
  • Δ mP ≥ 95 mP at 10% conversion of 10 μM ATP
  • Z’ and Δ mP specifications to be met using Transcreener ADP2 Assay reagents
  • Read time to achieve Z’ and Δ mP specifications ≤ 5 minutes

Look for these stickers on instruments that have successfully met the validation criteria.