PolarScreen™ Vitamin D Receptor Competitor Assay Kit, Red
PolarScreen™ Vitamin D Receptor Competitor Assay Kit, Red
Thermo Scientific™

PolarScreen™ Vitamin D Receptor Competitor Assay Kit, Red

The PolarScreen™ Vitamin D Receptor Competitor Assay Kit, Red, provides a sensitive and efficient method for high-throughput, fluorescence polarization-based screeningRead more
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Catalog number A15907
Price (USD)
2,110.00
Each
Add to cart
Price (USD)
2,110.00
Each
Add to cart

The PolarScreen™ Vitamin D Receptor Competitor Assay Kit, Red, provides a sensitive and efficient method for high-throughput, fluorescence polarization-based screening of potential vitamin D receptor (VDR) ligands. The kit uses insect cell-expressed, human full-length VDR (also available separately) and a novel, high-affinity, fluorescent VDR ligand (Fluormone™ VDR Red) in a homogenous, mix-and-read assay format.

This kit produces a red color read-out and is suitable to determine the IC50s of competitive compounds. For additional information or assistance contact us at drugdiscoverytech@lifetech.com.

The PolarScreen Vitamine D 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

VDR is added to a fluorescent VDR ligand (Fluormone VDR Red) to form a receptor/tracer complex, resulting in a high polarization value. This complex is then added to individual test compounds in microwell plates. Competitors displace the tracer from the complex, causing the fluorescent ligand to tumble more rapidly during its fluorescence lifetime, resulting in a low polarization value. Noncompetitors will not displace the fluorescent ligand from the complex; therefore, the polarization value will remain high. The shift in polarization value in the presence of test compounds is used to determine relative affinity of test compounds for VDR (see figure).

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
LigandVDR
Product TypeVitamin D Receptor Competitor Assay Kit
No. of Assays800 x 20 μL Assays
Unit SizeEach
Contents & Storage
• VDR Recombinant Human Protein, 1 tube (store at -68°C to -85°C)
• Fluormone VDR Red Tracer, 1 tube (-5°C to -30°C)
• VDR Red Screening Buffer, 3 bottles (-5°C to -30°C)
• DTT, 1 tube (-5°C to -30°C)

Frequently asked questions (FAQs)

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.