Targeting LRRK2 for Parkinson’s Disease with the Transcreener® GDP GTPase Assay

The Only Mix-and-Read Fluorescent GTPase Assay

The Transcreener GDP GTPase assay is a homogenous, fluorescent immunoassay that monitors the activity of GTPases along with GEF and GAP partners. The assay directly measures the amount of GDP produced in an in vitro GTPase reaction.

Learn More About Measuring GEF and GAP Activity:

Three Readouts Available

Three available fluorescent readouts provide a safe, HTS-compatible alternative to cumbersome radioassay methods and are more sensitive and less subject to interference than colorimetric phosphate detection methods such as malachite green. All readouts use far-red tracers to minimize compound interference and lower false-positive rates.

  • Fluorescence polarization (FP)
  • Fluorescence intensity (FI)
  • Time-resolved Forster-resonance-energy-transfer (TR-FRET)

GTPase Assay Principle - Universal GDP Detection

GTP hydrolysis from GTPase activity creates the product GDP and an additional phosphate. GDP displaces tracer from a highly selective monoclonal antibody resulting in a decrease in fluorescence polarization and TR-FRET or an increase in fluorescence intensity - perfect for your high throughput screen!

FP GTPase Assay Graphic
FI GTPase Assay Graphic
TR-FRET GTPase Graphic

What Can You Do With Transcreener

  • Measure guanine nucleotide exchange factor (GEF) activity
  • Measure GTPase-activating proteins (GAP) activity
  • Detect any GDP producing enzyme: GTPases, Gα proteins, Ras-like G proteins (Ras, Roc, Rac, cdc42, etc.), fucosyltransferases, mannosyltransferases

Assay Features

  • Easy-to-Use, Ultra-Sensitive GTPase Activity Assay
  • Homogenous, simple, mix-and-read method
  • Tunable dynamic range to match the target of interest
  • HTS Compatible with 96, 384, and 1536-well plates
  • Direct detection of GDP to monitor GTPase, GAP, and GEF Activity
  • Stable assay reagents – minimum 8-hour reagent and signal stability even at room temperature!

Single Step - Mix-and-Read

GDP GTPase Assay Schematic

Turnkey Protocol: Run enzyme reaction, add detection reagents, and read plates

Assay Tunable to Your Target

GTPase Standard Curve in GTP Hydrolysis

High Quality Data Under Initial Velocity Conditions

GTPase Assay Data Under Initial Velocity Conditions

Standard curves mimicking the enzymatic conversion of GTP to GDP using the Transcreener GDP FP Assay. The dynamic range of the assay can be easily tuned for different initial GTP concentrations. Excellent Z' values are achieved even at low GDP reagent formation.

Use with a Variety of GTPase Enzymes

Based on BellBrook Lab's proprietary nucleotide immunodetection technology, the Transcreener GDP Assays are compatible with any enzyme class that produces GDP, including monomeric small G proteins, such as CDC42, and Gα subunits of heterotrimeric G proteins, such as Gαi1. Learn how Transcreener can be used to study targets like Cdc42 and RhoA in Springer's Rho GTPases Methods and Protocols. Discover how researchers at UT Southwestern used the GDP assay to screen dynamin GTPase activity.

GTPase Enzyme Titration with Cdc42 and RhoA

Cdc42 and RhoA GTPase titration using the Transcreener GDP Assay with an FP readout.

GTPase Profiling Services

Interested in moving your program forward, but don't want to bring an assay in-house? Our scientists can help! BellBrook scientists right here in Madison, Wisconsin will use their extensive biochemistry and enzymology expertise to work with you and accelerate your GTPase program. We can provide inhibitor potency profiling for a variety of GTPase enzymes and related proteins. Get accurate IC50 results fast, to understand how your inhibitor interacts with other GTPases.

Assay Development Services

Contact us today to see if BellBrooks's GTPase profiling services will advance your research.

Far Red FP, FI &TR-FRET Readouts Validated on Major Multimode Readers

Supplier Instrument FP Assays
FI Assays
TR-FRET Assays
berthold logo TriStar²S LB 942 in review validated validated
Mithras² LB 943 in review validated validated
bioteklogo Cytation™ 5 validated validated validated
Cytation™ 3 validated validated validated
Cytation™ 1 validated validated validated
Synergy™ H1 validated validated validated
Synergy™ 2/H4/4 validated validated validated
Synergy™ HTX not capable validated not capable
Synergy™ Neo 2 validated validated validated
BMGLABTECH Logo POLARstar® Omega validated validated validated
FLUOstar® Omega not capable validated validated
PHERAstar® FSX validated validated validated
PHERAstar® Plus/FS validated validated validated
CLARIOstar® /Plus validated validated validated
VANTAstar validated validated validated
hidex logo Sense in review validated in review
MDS AT logo Analyst® GT/HT validated validated validated
Gemini® XPS/EM not capable validated not capable
SpectraMax® M2/M2e not capable validated not capable
SpectraMax® M5/M5e/FlexStation® 3 not capable validated validated
SpectraMax® Paradigm validated contact us contact us
SpectraMax® iD3/iD5 in review validated validated
perkinElmerLogo EnVision®/EnVision® Xcite validated validated validated
tecanLogo Infinite® M1000/M1000Pro/Safire2™ validated validated validated
Infinite® M200 not capable validated not capable
Infinite® F500 validated validated validated
Infinite® F200/Ultraevolution contact us validated contact us
Spark™ 10M validated validated validated