Predictor™ hERG Fluorescence Polarization Assay Kit

Zitierungen und Referenzen (1)

Predictor™ hERG Fluorescence Polarization Assay Kit

Katalognummer	Menge
PV5365	1 Kit

Katalognummer PV5365

Preis (EUR)

1.384,00

Zum Warenkorb hinzufügen

Menge:

1 Kit

Preis (EUR)

1.384,00

Zum Warenkorb hinzufügen

Das Predictor™ Assay-Kit für hERG-Fluoreszenzpolarisation enthält eine Reihe validierter Komponenten für die Durchführung von biochemischen hERG-Kanal-Bindungsstudien ohne Radioliganden. Der Assay wurde entwickelt, um potenzielle HERG-Kanalblockierer zu identifizieren und liefert Daten, die genau mit solchen aus elektrophysiologischen Patch-Clamp-Studien korrelieren. Der Assay basiert auf dem Prinzip der Fluoreszenzpolarisation, bei der eine rot-verschobenene fluoreszierende Markierungssubstanz durch Verbindungen, die an den hERG-Kanal binden, vom Kanal verdrängt wird. Das Leistungsverhalten des Assays wurde anhand einer Auswahl bekannter hERG-Kanalblocker validiert. Die Ergebnisse des Predictor™ Assays weisen eine hohe Korrelation mit den Ergebnissen von Patch-Clamp-Verfahren auf.

Nur für Forschungszwecke. Nicht zur Verwendung bei diagnostischen Verfahren.

Specifications

Assay-EingabeBiochemische Bindung

FarbeRot

NachweisverfahrenFluoreszent

Zielstruktur für MedikamentIonenkanäle

Zur Verwendung mit (Geräte)Mikrotiterplatten-Lesegerät

GensymbolhERG

Verpackung384-Well-Platte

ProdukttypAssay-Kit

ProteinfamilieIonenkanäle

Menge1 Kit

AnzeigewertEndpunkt

VersandbedingungTrockeneis

SystemtypPredictor™ System

ZieleintraghERG

KonjugatPredictor hERG Tracer rot

ProduktliniePredictor

Unit SizeEach

Inhalt und Lagerung

PV5365 Predictor™ hERG Fluoreszenzpolarisations-Assay-Kit
(andere Materialien müssen separat erworben werden; siehe Protokoll)
Lieferumfang:
• Predictor™ hERG Membranen
• Predictor™ hERG Tracer rot
• Predictor™ hERG Puffer
• E-4031
• Predictor™ hERG Fluoreszenzpolarisations-Assay-Protokoll
• Analysenzertifikat
Die Predictor™ hERG Membranen bei -80 °C lagern. Der Predictor™ Tracer rot, Predictor™ Puffer und E-4301 sollten bei -20 °C gelagert werden.

Häufig gestellte Fragen (FAQ)

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.

Zitierungen und Referenzen (1)

Zitierungen und Referenzen

Abstract

Development of the predictor HERG fluorescence polarization assay using a membrane protein enrichment approach.

Authors:Piper DR, Duff SR, Eliason HC, Frazee WJ, Frey EA, Fuerstenau-Sharp M, Jachec C, Marks BD, Pollok BA, Shekhani MS, Thompson DV, Whitney P, Vogel KW, Hess SD,

Journal:Assay Drug Dev Technol

PubMed ID:18471075

The life-threatening consequences of acquired, or drug-induced, long QT syndrome due to block of the human ether-a-go-go-related gene (hERG) channel are well appreciated and have been the cause of several drugs being removed from the market in recent years because of patient death. In the last decade, the propensity for ... More