Predictor™ hERG Fluorescence Polarization Assay Kit

Citas y referencias (1)

Predictor™ hERG Fluorescence Polarization Assay Kit

Kit de ensayo de polarización de la fluorescencia Predictor™ hERG

Número de catálogo	Cantidad
PV5365	1 Kit

Número de catálogo PV5365

Precio (EUR)

1.384,00

Cantidad:

1 Kit

Precio (EUR)

1.384,00

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.

El kit de ensayo de polarización de la fluorescencia Predictor™ hERG proporciona un conjunto de componentes validados para realizar estudios de unión bioquímica del canal hERG en ausencia de radioligandos. El ensayo se diseñó para identificar posibles bloqueadores del canal hERG mediante la producción de datos que se correlacionan con precisión con estudios de electrofisiología con pinzamiento zonal. El ensayo se basa en el principio de polarización de la fluorescencia, donde un marcador rojo fluorescente de desplazamiento se desplaza del canal hERG por compuestos que se unen al canal. El rendimiento del ensayo se validó con un panel de bloqueadores de canal hERG establecidos. Los resultados del ensayo Predictor™ demuestran una alta correlación con los obtenidos de las técnicas de pinzamiento zonal.

Para uso exclusivo en investigación. No apto para uso en procedimientos diagnósticos.

Especificaciones

Entrada del ensayoUnión bioquímica

ColorRojo

Método de detecciónFluorescente

Diana del fármacoCanales de iones

Para utilizar con (equipo)Lector de microplacas

Símbolo de genhERG

EnvasePlaca de 384 pocillos

Tipo de productoKit de ensayo

Familia de proteínasCanales de iones

Cantidad1 Kit

Presentación de datosPunto final

Condiciones de envíoHielo seco

Tipo de sistemaSistema Predictor™

Entrada dianahERG

ConjugadoMarcador rojo Predictor hERG

Línea de productosPredictor

Unit SizeEach

Contenido y almacenamiento

Kit de ensayo de polarización de la fluorescencia PV5365 Predictor™ hERG
(Otros materiales se solicitan por separado; consulte el protocolo)
Incluye:
• Membranas Predictor™ hERG
• Marcador rojo Predictor™ hERG
• Tampones Predictor™ HERG
• E-4031
• Protocolo de ensayo de polarización de la fluorescencia Predictor™ hERG
• Certificado de Análisis
Almacene las membranas Predictor™ hERG a -80 °C. El marcador rojo Predictor™, los tampones Predictor™ y E-4301 deben almacenarse a -20 °C.

Preguntas frecuentes

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.

Citations & References (1)

Citations & References

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