PolarScreen™ Glucocorticoid Receptor Competitor Assay Kit, Red
PolarScreen™ Glucocorticoid Receptor Competitor Assay Kit, Red

PolarScreen™ Glucocorticoid Receptor Competitor Assay Kit, Red

The PolarScreen™ Glucocorticoid Receptor Competitor Assay Kit, Red, provides a sensitive and efficient method for high-throughput, fluorescence polarization-based screening ofRead more
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Catalog number A15898
Price (USD)
1,312.00
Each
Add to cart
Price (USD)
1,312.00
Each
Add to cart
The PolarScreen™ Glucocorticoid Receptor Competitor Assay Kit, Red, provides a sensitive and efficient method for high-throughput, fluorescence polarization-based screening of potential glucocorticoid receptor (GR) ligands. The kit uses insect cell-expressed, human GR (also available separately) and a novel, high-affinity, fluorescent glucocorticoid ligand (Fluormone™ GS Red) in a homogenous, mix-and-read assay format.

This kit produces a red color read-out; a 'green' kit is also available. 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, contact drugdiscoverytech@lifetech.com.

The PolarScreen Glucocorticoid Receptor Competitor Assay is:

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

How the Assay Works
GR is added to a fluorescent glucocorticoid ligand (Fluormone GS Red) in the presence of competitor test compounds in microwell plates. The presence of effective competitors prevents the formation of a GS Red/GR complex, resulting in a decrease of the polarization value. The shift in polarization value in the presence of test compounds is used to determine relative affinity of test compounds for GR. The assay is designed to be carried out at room temperature through use of a coactivator-related stabilizing peptide. Alternatively, the assay may be run at 4°C in the absence of the stabilizing peptide, with insignificant change in GR binding characteristics.

Contact us for bulk quantities or for assistance with other nuclear receptor assay needs, from assay development to library screening and beyond.
WARNING: Reproductive Harm - www.P65Warnings.ca.gov
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Product LinePolarScreen™
For Use With (Application)Competitive Binding Assay, Nuclear Receptor Assay
LigandGR
Product TypeAssay Kit
No. of Assays400 x 20 μL assays
Unit SizeEach
Contents & Storage
1 tube Glucocorticoid Receptor Recombinant Protein (store at -68 to -85°C)
1 tube of Fluormone GS Red Tracer (store at -5 to -30°C)
3 tubes of GR Screening Buffer (store at -5 to -30°C)
3 tubes of GR Stabilizing Peptide (store at -68 to -85°C)
1 tube of DTT (store at -5 to -30°C)

Frequently asked questions (FAQs)

How do I export Ct values in the TaqMan hPSC Scorecard Analysis Software?

The only way to view the Ct values in the current version is through the expression plot. We expect the software's Excel export to include the gene names and Ct values in a future version of the software.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Applications 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.