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Predictor™ hERG Fluorescence Polarization Assay Kit
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Predictor™ hERG Fluorescence Polarization Assay Kit

Predictor™ hERG Fluorescence Polarization Assay Kit

The Predictor™ hERG Fluorescence Polarization Assay Kit provides a set of validated components to perform hERG channel biochemical binding studiesRead more
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Catalog NumberQuantity
PV53651 Kit
Catalog number PV5365
Price (USD)
1,458.00
Each
Add to cart
Quantity:
1 Kit
Price (USD)
1,458.00
Each
Add to cart
The Predictor™ hERG Fluorescence Polarization Assay Kit provides a set of validated components to perform hERG channel biochemical binding studies in the absence of radioligand. The assay was designed to identify potential hERG channel blockers by producing data that accurately correlates with patch-clamp electrophysiology studies. The assay is based on the principle of fluorescence polarization where a red-shifted fluorescent tracer is displaced from the hERG channel by compounds that bind to the channel. Assay performance was validated with a panel of established hERG channel blockers. Results from the Predictor™ assay demonstrate a high correlation with those obtained from patch clamp techniques.
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Assay EntryBiochemical binding
ColorRed
Detection MethodFluorescence
Druggable TargetIon Channels
For Use With (Equipment)Microplate Reader
Gene SymbolhERG
Packaging384-well Plate
Product TypeAssay Kit
Protein FamilyIon Channels
Quantity1 Kit
ReadoutEnd Point
Shipping ConditionDry Ice
System TypePredictor™ System
Target EntryhERG
ConjugatePredictor hERG Tracer Red
Product LinePredictor™
Unit SizeEach
Contents & Storage

PV5365 Predictor™ hERG Fluorescence Polarization Assay Kit
(Other materials are required separately; please refer to the protocol)
Includes:
• Predictor™ hERG Membranes
• Predictor™ hERG Tracer Red
• Predictor™ hERG Buffers
• E-4031
• Predictor™ hERG Fluorescence Polarization Assay Protocol
• Certificate of Analysis
Store the Predictor™ hERG Membranes at -80°C. The Predictor™ Tracer Red, Predictor™buffers, and E-4301 should be stored at -20°C.

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Frequently asked questions (FAQs)

I am doing drug discovery. Can I use the Predictor hERG Membranes for radioligand binding?

No, the Predictor hERG Membranes are not suitable for radioligand binding.
These membranes contain extremely high levels of hERG channel protein. Radioligand binding assays on these membranes would require either abnormally high concentrations of radioligand or severe dilutions of the membrane preparation, which tend to have a deleterious effect on hERG channel conformation and stability.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

What are the common issues encountered with FP assays?

-No assay window: Check that your instrument is designed to do FP experiments.
-Poor discrimination between the minimum and maximum mP Control (both include the receptor): This may be an indication of the quality of the receptor. Do not make single use aliquots of the receptor or store diluted.
-The type of microplate is critical: Please see if the protocol specifies untreated polystyrene or NBS-coated plates. If the plate binds the fluorescent substrate, it cannot freely rotate and the mP reading will always be high. White plates cannot be used for FP assays. Avoid the use of silanized pipette tips and vials.
-The “No Receptor Control' (free Fluormone/Tracer) should have much higher RFU values compared to buffer alone. If not, there is a problem with the instrument setup or less commonly with the Fluormone/Tracer.
-If the free Fluormone/Tracer mP value is greater than 100 mP or negative, please refer to the user manual for your instrument and reset the G factor.

Find additional tips, troubleshooting help, and resources within our Drug Discovery & Development Support Center.

What is the best type of microplate reader for fluorescence polarization (FP) assays?

For FP assays, plate readers must specifically have FP built in. Please see our instrument compatibility portal (http://www.thermofisher.com/us/en/home/industrial/pharma-biopharma/drug-discovery-development/dd-misc/instrument-compatibility-portal.html).

Find additional tips, troubleshooting help, and resources within our Drug Discovery & Development Support Center.

How can I test the fluorescence polarization (FP) settings of my microplate reader before doing the assay?

We offer a FP One-Step Reference Kit (https://www.thermofisher.com/order/catalog/product/P3088) to detect in green (Ex/Em = 488/535 nm) and red (Ex/Em = 525/590 nm) emission ranges, suitable for the PolarScreen Green and Red Assays. The alternative is to perform the Controls in the respective assay kit prior to performing the actual assay using test compounds.

Find additional tips, troubleshooting help, and resources within our Drug Discovery & Development Support Center.

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