BellBrook

Transcreener GDP FP Assay

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BBL-3009-1K1000 assaysCHF 771.00
BBL-3009-10K10000 assaysCHF 3'293.00
BBL-3009-100K10 x 10000 assaysINQ
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Product Details
Product Type Kit
Properties
Application Set Compound Screening
Crossreactivity All
Quantity

1000 assays/384wells (BBL-3009-1K)

10000 assays/384wells (BBL-3009-10K)

Note: The exact number of assays depends on enzyme reaction conditions.

Sensitivity Excellent data quality (Z’ ≥ 0.7) at low substrate conversion (typically 10–30%).
Range Accommodates GTP concentrations ranging from 1 μM to 1,000 μM.
Detection Type Fluorescent polarization
Kit Contains

GDP Antibody, GDP Alexa Fluor® 633 Tracer, Stop & Detect Buffer B, 10X, GTP, GDP

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
Our product description may differ slightly from the original manufacturers product datasheet.


Assay Features

Easy-to-Use (mix-and-read method), ultra-sensitive GTPase activity assay. HTS Compatible with 96, 384, and 1536-well plates. Tunable dynamic range to match the target of interest. Stable assay reagents - minimum 8-hour reagent and signal stability even at room temperature! Direct detection of GDP!

Declaration Manufactured by BellBrook Labs.
Shipping and Handling
Shipping DRY ICE
Short Term Storage -20°C
Long Term Storage -20°C
Handling Advice Avoid freeze/thaw cycles.
Documents
Manual Download PDF
MSDS Inquire
Product Specification Sheet
Datasheet Download PDF
Description

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 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. Three available fluorescent readouts, Fluorescence polarization (FP), Fluorescence intensity (FI) and Time-resolved Forster-resonance-energy-transfer (TR-FRET), provide a safe HTS-compatible alternative to cumbersome radioassay methods and are more sensitive and less subject to interference than colorimetric phosphate detection methods. All readouts use far-red tracers to minimize compound interference and lower false-positive rates.

Transcreener® GDP FP Assay (Prod. No. BBL-3009) is a mix-and-read fluorescent GTPase assay to follow the progress of any enzyme that produces GDP, including monomeric small G proteins, such as CDC42, and Gα subunits of heterotrimeric G proteins, such as Gαi1. It can be used for screening GTPases, fucosyltransferase or mannosyltransferase and covers a broad range of GTP concentrations. Highly selective, high-affinity antibody enables nanomolar GDP detection in the presence of excess GTP. This assay accommodates GTP concentrations ranging from 1 μM to 1,000 μM with excellent data quality (Z’ ≥ 0.7) at low substrate conversion (typically 10-30%).

Product References
  1. Two Galpha(i1) rate-modifying mutations act in concert to allow receptor-independent, steady-state measurements of RGS protein activity: T. Zielinski, et al.; J. Biomol. Screen. 14, 1195 (2009)
  2. High-throughput fluorescence polarization assay for the enzymatic activity of GTPase-activating protein of ADP-ribosylation factor (ARFGAP): W. Sun, et al.; J. Biomol. Screen. 16, 717 (2011)
  3. Evaluating modulators of "Regulator of G-protein Signaling" (RGS) proteins: D.E. Bosch, et al.; Curr. Protoc. Pharmacol. Chapter 2 Unit 2.8 (2012)
  4. A High-Throughput Assay for Rho Guanine Nucleotide Exchange Factors Based on the Transcreener GDP Assay: M. Reichman, et al.; J. Biomol. Screen. 20, 1294 (2015)
  5. A highly-sensitive high throughput assay for dynamin's basal GTPase activity: A. Mohanakrishnan, et al.; PLoS One 12, e0185639 (2017)
  6. High-Throughput Assay for RhoGEFs Based on the Transcreener® GDP Assay: R.G. Lowery & M. Kumar; Meth. Mol. Biol. 1821, 165 (2018)
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