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  5. Invitrogen™ PolarScreen™ ER Beta Competitor Assay Kit, Green

Invitrogen™ PolarScreen™ ER Beta Competitor Assay Kit, Green

Catalog No. A15890
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A15890 800 Assays
A15889 2500 Assays
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Description

The PolarScreen™ ER Beta Competitor Assay Kit provides a sensitive and efficient method for high-throughput, fluorescence polarization-based screening of potential estrogen receptor (ER) beta ligands.

The PolarScreen™ ER Beta Competitor Assay Kit provides a sensitive and efficient method for high-throughput, fluorescence polarization-based screening of potential estrogen receptor (ER) beta ligands. The kit uses insect cell-expressed, full-length, native (untagged) human ER beta (Cat. No. A15664) and a novel, high-affinity, fluorescent estrogen ligand (Fluormone™ ES2) in a homogenous, mix-and-read assay format.

This kit produces a green color read-out; a 'red' kit is also available (A15891). Both kits suitably determine the IC50s of competitive compounds. Color choice depends on the spectral properties of the compounds to be screened and the available instrumentation. For additional information or assistance email drugdiscoverytech@lifetech.com.

The PolarScreen™ ER Beta Competitor Assay Kit is:

  • Complete & ready-to-use out of the box—all reagents included, just add your test compounds
  • High throughput compatible—20 μL fluorescence polarization assays in a 384-well plate
  • Convenient—available in bulk sizes for larger screening needs

How the Assay Works

Full-length ER beta is added to a fluorescent estrogen ligand to form an ER-Fluormone™ ES2 complex. This complex is added to individual test compounds in multiwell plates. To determine relative affinity, the assay measures fluorescence polarization. Competing test compounds will displace the Fluormone™ ES2 ligand from ER beta, permitting it to tumble rapidly and resulting in low polarization values. A shift in the fluorescence polarization value in the presence of test compound is used to determine the relative affinity of test compounds for ER beta.

All components are manufactured under the strictest quality parameters. This product's performance, per the Certificat of Analysis, is guaranteed for 6 months after purchase. Contact us for bulk quantities or for assistance with other nuclear receptor assay needs, from assay development to library screening and beyond.

Specifications

Color Green
Quantity 800 Assays
Packaging 384-well Plate
Content And Storage • Estrogen Receptor (ER) Beta Recombinant Protein, Full Length (store at -68°C to -85°C)
• Fluromone™ ES2 (store at -5°C to -30° C)
• ES2 Screening Buffer (store at room temperature)
Assay Category Biochemical Assay
Druggable Target Nuclear Receptors
Readout Fluorescence Polarization
Product Line PolarScreen
For Use With (Application) Nuclear Receptor Assay, Fluorescence Polarization
Ligand ER beta
Product Type Assay Kit
Target Molecule Estrogen Receptor Beta Nuclear Receptor
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Frequently Asked Questions (FAQs)

Where do I find more information about how to use the TaqMan hPSC Scorecard Analysis Software?

The link for the software user guide can be found on this page.
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.
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).
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.

For Research Use Only. Not for use in diagnostic procedures.