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Invitrogen™ PolarScreen™ Progesterone Receptor Competitor Assay Kit, Green

Catalog No. A15905
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400 x 20 μL assays
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A15905 400 x 20 μL assays
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Catalog No. A15905 Supplier Invitrogen™ Supplier No. A15905
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Description

PolarScreen™ Progesterone Receptor Competitor Assay Kit, Green

The PolarScreen™ Progesterone Receptor Competitor Assay Kit, Green, provides a sensitive and efficient method for high-throughput, fluorescence polarization-based screening of potential progesterone receptor (PR) ligands. The kit uses insect cell-expressed, human PR ligand binding domain tagged with glutathione-S-transferase (GST) (also available separately) and a novel, high-affinity, fluorescent progesterone receptor ligand (Fluormone™ PL Green) in a homogenous, mix-and-read assay format.

This kit produces a green color read-out; a 'red' 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 us at drugdiscoverytech@lifetech.com.

The PolarScreen Progesterone 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
    An N-terminal fusion of glutathione transferase to the ligand-binding domain of the human progesterone receptor (PR-LBD(GST)) is added to a fluorescently-tagged progesterone ligand (Fluormone PL Green) in the presence of competitor test compounds in microwell plates. The presence of effective competitors prevents the formation of a PL Green/PR-LBD(GST) 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 PR-LBD(GST).

    Contact us for bulk quantities or for assistance with other nuclear receptor assay needs, from assay development to library screening and beyond.

    • Specifications

    Quantity 400 x 20 μL assays
    Content And Storage 1 tube Progesterone Receptor Ligand Binding Domain Recombinant Protein (store at -68 to -85°C)
    1 tube of Fluormone PL Green (store at -5 to -30°C)
    2 bottles of PR Screening Buffer (store at -5 to -30°C)
    1 tube of 1 M DTT (store at -5 to -30°C)
    Product Line PolarScreen
    For Use With (Application) Competitive Binding Assay, Nuclear Receptor Assay
    Ligand PR
    Product Type Assay Kit
    No. of Assays 400 x 20 μL assays

    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.
    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.