Biochemical Nuclear Receptor Assays - Kits

PolarScreen™ Androgen Receptor Competitor Assay Kit, Red

The PolarScreen™ Androgen Receptor Competitor Assay Kit, Red, provides a sensitive and efficient method for high-throughput, fluorescence polarization-based screening of potential androgen receptor (AR) ligands. The kit uses insect cell-expressed rat AR ligand binding domain tagged with glutathione-S-transferase (GST) and histidine (also available separately), plus a novel, high-affinity, fluorescent androgen receptor ligand (Fluormone AL 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 us at drugdiscoverytech@lifetech.com.

The PolarScreen Androgen 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
The kit uses the rat AR ligand-binding domain tagged with His and GST [AR-LBD(His-GST)]. AR-LBD(His-GST) is added to a fluorescently-tagged androgen ligand (FluormoneAL Red) in the presence of competitor test compounds in microwell plates. Competitors displace the fluorescent FluormoneAL Red Ligand from the AR-LBD/FluormoneAL Red complex, causing the fluorescent ligand to tumble 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 remains high. The shift in polarization value in the presence of the test compounds is used to determine relative affinity of test compounds for the AR-LBD.

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

LanthaScreen™ TR-FRET TR beta Coactivator Assay Kit, goat

This kit contains Goat Tb-Anti-GST antibody; other kit components are the same as kit A15135:

The LanthaScreen® TR-FRET TR (thyroid receptor) beta Coactivator Assay Kit provides a sensitive and robust method for high-throughput screening of potential TR beta ligands as agonists of ligand-dependent coactivator recruitment. The kit uses a terbium (Tb)-labeled anti-GST antibody, a fluorescein-labeled coactivator peptide, and a human recombinant TR beta ligand-binding domain (TR beta-LBD) that is tagged with glutathione-S-transferase (GST) in a homogeneous, mix-and-read assay format.

To Assay:
When performing the LanthaScreen® TR-FRET TR beta coactivator assay, TR beta-LBD is added to ligand test compounds followed by addition of a mixture of the fluorescein-coactivator peptide and Tb anti-GST antibody. After an incubation period at room temperature, the 520 nm/495 nm TR-FRET ratio is calculated and can be used to determine the EC50 from a dose response curve of the compound. Based on the biology of the TR beta-coactivator peptide interaction, the ligand EC50 is a composite value representing the amount of ligand required to bind to receptor, induce a conformational change, and recruit coactivator peptide (Figure 1).

Contents and Storage:
The LanthaScreen® TR-FRET TR beta Coactivator Assay Kit contains TR beta-LBD (GST) protein, fluorescently labeled SRC2-2 coactivator peptide, Tb-labeled anti-GST antibody, and buffers. Store components as indicated in the assay protocol (-80°C, -20°C, or +4°C).

LanthaScreen™ TR-FRET ERR gamma Coactivator Assay Kit, goat

This kit contains Goat Tb-Anti-GST antibody; other kit components are the same as kit A15121:

The LanthaScreen® TR-FRET Estrogen Related Receptor (ERR) gamma Coactivator Assay Kit provides a sensitive and robust method for high-throughput screening of potential ERR gamma ligands as antagonists/inverse agonists of coactivator displacement. The homogeneous mix-and-read assay format results in a ligand EC50 composite value representing the amount of ligand required to bind to receptor, effect a conformational change, and displace coactivator peptide (Figure 1).

How it works

The LanthaScreen® TR-FRET ERR gamma Coactivator Assay Kit includes a terbium (Tb)-labeled anti-GST antibody, a fluorescein-labeled coactivator peptide, and a human ERR gamma ligand-binding domain (ERR gamma-LBD) that is tagged with glutathione-S-transferase (GST). To assay, ERR gamma-LBD is added to test compounds, followed by addition of a mixture of the fluorescein-coactivator peptide and Tb-anti- GST antibody. After room temperature incubation, the TR-FRET 520:495 emission ratio is calculated and used to determine the EC50 from a dose response curve of the compound.

Contents and Storage:

The LanthaScreen® TR-FRET ERR gamma Coactivator Assay Kit contains ERR gamma-LBD (GST) protein, fluorescently labeled PGC1a coactivator peptide, Tb-anti-GST antibody, and buffers. Store components as indicated in the assay protocol (-80°C, -20°C, or +4°C).

LanthaScreen™ TR-FRET LXR beta Coactivator Assay Kit, goat

This kit contains Goat Tb-Anti-GST antibody; other kit components are the same as kit A15129:

The LanthaScreen® TR-FRET LXR (liver X receptor) beta Coactivator Assay provides a sensitive and robust method for high-throughput screening of potential LXR beta ligands as agonists of coactivator recruitment. The kit uses a terbium (Tb)-labeled anti-GST antibody, a fluorescein-labeled coactivator peptide, and an LXR beta ligand-binding domain (LBD) that is tagged with glutathione-S-transferase (GST), in a homogeneous mix-and-read assay format.

How it works:
In the LanthaScreen® TR-FRET LXR beta Coactivator Assay, LXR beta-LBD is added to ligand test compounds followed by the addition of a mixture of the fluorescein-coactivator peptide and Tb-labeled anti-GST antibody. After an incubation period at room temperature, the 520 nm/495 nm TR-FRET ratio is calculated and can be used to determine the EC50 from a dose response curve of the compound. Based on the biology of the LXR beta-coactivator peptide interaction, this ligand EC50 is a composite value representing the amount of ligand required to bind to receptor, effect a conformational change, and recruit coactivator peptide (Figure 1).

Contents and Storage:
The LanthaScreen® TR-FRET LXR beta Coactivator Assay Kit contains LXR beta-LBD (GST) protein, fluorescently labeled D22 coactivator peptide, Tb-labeled anti-GST antibody, and buffers. Store components as indicated in the assay protocol (-80°C, -20°C, or +4°C).

PolarScreen™ ER Alpha Competitor Assay, Green

The PolarScreen™ ER (Estrogen Receptor) Alpha Competitor Assay Kit, Green, provides a sensitive and efficient method for high-throughput, fluorescence polarization-based screening of potential ER alpha ligands. The kit uses insect cell-expressed, full-length, native (untagged) human estrogen receptor alpha (available separately, Cat. No. A15674) and a novel, high-affinity, fluorescent estrogen ligand (Fluormone ES2 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 drugdiscoverytech@lifetech.com.

PolarScreen™ ER Alpha Competitor Assay, Green, is:

• Complete & ready-to-use out of the box—all reagents included, just add your test compounds
• High throughput compatible—800 x 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 alpha 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 alpha, 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 alpha.

All components are manufactured under the strictest quality parameters. This product's performance, per the Certificate 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.

PolarScreen™ PPARγ-Competitor Assay Kit, Green

The PPARγ (Peroxisome Proliferator-Activated Receptor-gamma) Competitor Assay, Green uses the human-derived recombinant PPARγ ligand-binding domain (PPARγ-LBD) tagged with an N-terminal GST-tag and a novel, tight-binding, selective fluorescent PPARγ ligand (Fluormone™ PPARγ Green) in a homogenous, mix-and-read assay format. PPARγ-LBD/Fluormone™ PPARγ Green complex produces high polarization. This complex is then added to individual test compounds in microwell plates. Competitors displace the fluorescent Fluormone™ PPARγ Green Ligand from the PPARγ-LBD/Fluormone™ PPARγ Green complex, causing the fluorescent ligand to tumble rapidly during its fluorescence lifetime, resulting in a low polarization value. Noncompetitors will not displace the fluorescent ligand from the complex, so the polarization value remains high. The shift in polarization value in the presence of the test compounds is used to determine relative affinity of test compounds for the PPARγ-LBD.

LanthaScreen™ TR-FRET ER beta Coactivator Assay Kit

The LanthaScreen® TR-FRET ER beta Coactivator Assay Kit provides a sensitive and robust method for high-throughput screening of potential estrogen receptor (ER) beta ligands as agonists or antagonists of ligand-dependent coactivator recruitment. The kit uses a ER beta ligand-binding domain (ER beta-LBD) that is tagged with glutathione-S-transferase (GST), a terbium (Tb)-labeled anti-GST antibody, and a fluorescein-labeled coactivator peptide in a homogenous mix-and-read assay format.

Agonist Mode
To run the LanthaScreen TR-FRET ER beta Coactivator Assay in agonist mode (to identify agonist compounds), ER beta-LBD is added to ligand test compounds followed by the addition of a mixture of fluorescein-coactivator peptide and Tb-anti-GST antibody. After room temperature incubation, the TR-FRET 520:495 emission ratio is calculated and used to determine the EC50 from a dose response curve of the compound. Based upon the biology of the ER beta-coactivator peptide interaction, this ligand EC50 is a composite value representing the amount of ligand required to bind to receptor, effect a conformational change, and recruit coactivator peptide.

Antagonist Mode
When the LanthaScreen TR-FRET ER beta Coactivator Assay Kit is run in antagonist mode (to identify antagonist compounds), ER beta-LBD is added to ligand test compounds followed by the addition of a mixture of agonist, fluorescein-coactivator peptide, and Tb-anti-GST antibody. The concentration of agonist used in this mode is the EC80 concentration as determined by first running the assay in agonist mode.

LanthaScreen™ TR-FRET PPAR alpha Coactivator Assay Kit, rabbit

This kit contains Rabbit Tb-Anti-GST antibody; other kit components are the same as kit PV4684:

The LanthaScreen® TR-FRET PPAR (peroxisome proliferator activated receptor) alpha Coactivator Assay provides a sensitive and robust method for high-throughput screening of potential PPAR alpha ligands as agonists or antagonists of ligand-dependent coactivator recruitment. The kit uses a terbium (Tb)-labeled anti-GST antibody, a fluorescein-labeled coactivator peptide, and a human recombinant PPAR alpha ligand-binding domain (PPAR alpha-LBD) that is tagged with glutathione-S-transferase (GST) in a homogeneous mix-and-read assay format.

Agonist mode:
When running the LanthaScreen® TR-FRET PPAR alpha Coactivator Assay in agonist mode (to identify agonist compounds), PPAR alpha- LBD is added to ligand test compounds followed by addition of a mixture of the fluorescein-coactivator peptide and Tb-labeled anti-GST antibody. After an incubation period at room temperature, the 520 nm/495 nm TR-FRET ratio is calculated and can be used to determine the EC50 from a dose response curve of the compound. Based on the biology of the PPAR alpha-coactivator peptide interaction, this ligand EC50 is a composite value representing the amount of ligand required to bind to receptor, effect a conformational change, and recruit coactivator peptide (see Figure 1).

Antagonist mode:
When the LanthaScreen® TR-FRET PPAR alpha Coactivator Assay is run in antagonist mode (to identify antagonist compounds), PPAR alpha-LBD is added to ligand test compounds followed by addition of a mixture of agonist, fluorescein-coactivator peptide, and Tb-labeled anti-GST antibody (Figure 2). The concentration of agonist used in this mode is the EC80 concentration as determined by first running the assay in agonist mode.

Contents and Storage:
The LanthaScreen® TR-FRET PPAR alpha Coactivator Assay Kit contains PPAR alpha-LBD (GST) protein, fluorescently labeled PGC1a coactivator peptide, Tb-labeled anti-GST antibody, and buffers. Store components as indicated in the assay protocol (-80°C, -20°C, or 4°C).

LanthaScreen™ TR-FRET TR alpha Coactivator Assay Kit, goat

This kit contains Goat Tb-Anti-GST antibody; other kit components are the same as kit A15136:

The LanthaScreen® TR-FRET TR (thyroid receptor) alpha Coactivator Assay Kit provides a sensitive and robust method for highthroughput screening of potential TR alpha ligands as agonists of ligand-dependent coactivator recruitment. The kit uses a terbium (Tb)-labeled anti-GST antibody, a fluorescein-labeled coactivator peptide, and a human recombinant TR alpha ligand-binding domain (TR alpha-LBD) that is tagged with glutathione-S-transferase (GST) in a homogeneous, mix-and-read assay format.

To Assay:
When performing the LanthaScreen® TR-FRET TR alpha coactivator assay, TR alpha-LBD is added to ligand test compounds followed by addition of a mixture of the fluorescein-coactivator peptide and Tb anti-GST antibody. After an incubation period at room temperature, the 520 nm/495 nm TR-FRET ratio is calculated and used to determine the EC50 from a dose response curve of the compound. Based on the biology of the TR alpha-coactivator peptide interaction, the ligand EC50 is a composite value representing the amount of ligand required to bind to receptor, induce a conformational change, and recruit coactivator peptide (Figure 1).

Contents and Storage:
The LanthaScreen® TR-FRET TR alpha Coactivator Assay Kit contains TR alpha-LBD (GST) protein, fluorescently labeled SRC2-2 coactivator peptide, Tb-labeled anti-GST antibody, and buffers. Store components as indicated in the assay protocol (-80°C, -20°C, or +4°C).

LanthaScreen™ TR-FRET RAR alpha Coactivator Assay Kit, goat

This kit contains Goat Tb-Anti-GST antibody; other kit components are the same as kit A15122:

The LanthaScreen® TR-FRET Retinoic Acid Receptor (RAR) alpha Coactivator Assay Kit provides a sensitive and robust method for high-throughput screening of potential RAR alpha ligands as agonists of coactivator recruitment. The homogeneous mix-and-read assay format results in a ligand EC50 composite value representing the amount of ligand required to bind to receptor, effect a conformational change, and recruit coactivator peptide (Figure 1).

How it works

The LanthaScreen® TR-FRET RAR alpha Coactivator Assay Kit includes a terbium (Tb)-labeled anti-GST antibody, a fluorescein-labeled coactivator peptide, and a human RAR alpha ligand-binding domain (RAR alpha-LBD) that is tagged with glutathione-S-transferase (GST). To assay, RAR alpha-LBD is added to ligand test compounds, followed by addition of a mixture of the fluorescein-coactivator peptide and Tb-anti-GST antibody. After room temperature incubation, the TR-FRET 520:495 emission ratio is calculated and used to determine the EC50 from a dose response curve of the compound.

Contents and Storage:

The LanthaScreen® TR-FRET RAR alpha Coactivator Assay Kit contains RAR alpha-LBD (GST) protein, fluorescein-labeled D22 coactivator peptide, Tb-anti-GST antibody, and buffers. Store components as indicated in the assay protocol (-80°C, -20°C, or +4°C).

LanthaScreen™ TR-FRET ER beta Competitive Binding Kit

The LanthaScreen® TR-FRET ER beta Competitive Binding Kit provides a sensitive and robust method for high-throughput screening (HTS) of ligands for estrogen receptor (ER) beta. The kit uses a human ER beta ligand-binding domain (LBD) that is tagged with glutathione S-transferase (GST), a terbium-labeled anti-GST antibody, and a fluorescent small-molecule ER ligand (Fluormone™ ES2) in a homogeneous mix-and-read assay format.

The Lanthascreen ER beta Competitive Binding Assay is:

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

LanthaScreen™ TR-FRET ERR gamma Coactivator Assay Kit, rabbit

This kit contains Rabbit Tb-Anti-GST antibody; other kit components are the same as kit PV4408:

The LanthaScreen® TR-FRET Estrogen Related Receptor (ERR) gamma Coactivator Assay Kit provides a sensitive and robust method for high-throughput screening of potential ERR gamma ligands as antagonists/inverse agonists of coactivator displacement. The homogeneous mix-and-read assay format results in a ligand EC50 composite value representing the amount of ligand required to bind to receptor, effect a conformational change, and displace coactivator peptide (Figure 1).

How it works

The LanthaScreen® TR-FRET ERR gamma Coactivator Assay Kit includes a terbium (Tb)-labeled anti-GST antibody, a fluorescein-labeled coactivator peptide, and a human ERR gamma ligand-binding domain (ERR gamma-LBD) that is tagged with glutathione-S-transferase (GST). To assay, ERR gamma-LBD is added to test compounds, followed by addition of a mixture of the fluorescein-coactivator peptide and Tb-anti- GST antibody. After room temperature incubation, the TR-FRET 520:495 emission ratio is calculated and used to determine the EC50 from a dose response curve of the compound.

Contents and Storage:

The LanthaScreen® TR-FRET ERR gamma Coactivator Assay Kit contains ERR gamma-LBD (GST) protein, fluorescently labeled PGC1a coactivator peptide, Tb-anti-GST antibody, and buffers. Store components as indicated in the assay protocol (-80°C, -20°C, or +4°C).

LanthaScreen™ TR-FRET ERR alpha Coactivator Assay Kit, goat

This kit contains Goat Tb-Anti-GST antibody; other kit components are the same as kit A15130:

The LanthaScreen® ERR (estrogen related receptor) alpha TR-FRET Coactivator Assay provides a sensitive and robust method for high-throughput screening of potential ERR alpha ligands as inverse agonists (sometimes referred to as antagonists) of coactivator displacement. The kit uses a terbium (Tb)-labeled anti-GST antibody, a fluorescein-labeled coactivator peptide, and an ERR alpha ligand-binding domain (ERR alpha-LBD) that is tagged with glutathione-S-transferase (GST), in a homogeneous mix-and-read assay format.


In the LanthaScreen® ERR alpha TR-FRET Coactivator Assay, ERR alpha-LBD is added to ligand test compounds, followed by addition of a mixture of the fluorescein-coactivator peptide and Tb-labeled anti-GST antibody. After an incubation period at room temperature, the 520 nm/495 nm TR-FRET ratio is calculated and can be used to determine the EC50 from a dose response curve of the compound. Based on the biology of the ERR alpha-coactivator peptide interaction, this ligand EC50 is a composite value representing the amount of ligand required to bind to receptor, effect a conformational change, and displace coactivator peptide (Figure 1).


Contents and Storage:


The LanthaScreen® TR-FRET ERR alpha Coactivator Assay Kit contains ERR alpha-LBD (GST) protein, fluorescently labeled PGC1a coactivator peptide, Tb-labeled anti-GST antibody, and buffers. Store components as indicated in the assay protocol (-80°C, -20°C, or +4°C).

LanthaScreen™ TR-FRET FXR Coactivator Assay Kit, rabbit

This kit contains Rabbit Tb-Anti-GST antibody; other kit components are the same as kit PV4833:

The LanthaScreen® TR-FRET FXR (farnesoid X receptor) Coactivator Assay Kit provides a sensitive and robust method for high-throughput screening of potential FXR ligands as agonists of coactivator recruitment. The kit uses a terbium (Tb)-labeled anti-GST antibody, a fluorescein- labeled coactivator peptide, and an FXR ligand-binding domain (FXR-LBD) that is tagged with glutathione-S-transferase (GST), in a homogeneous, mix-and-read assay format.

To assay:
When running the LanthaScreen® TR-FRET FXR coactivator assay in agonist mode, FXR-LBD is added to ligand test compounds followed by addition of a mixture of the fluorescein-coactivator peptide and Tb-labeled anti-GST antibody. After an incubation period at room temperature, the 520 nm/495 nm TR-FRET ratio is calculated and used to determine the EC50 from a dose response curve of the compound. Based on the biology of the FXR-coactivator peptide interaction, the ligand EC50 is a composite value representing the amount of ligand required to bind to receptor, effect a conformational change, and recruit coactivator peptide (Figure 1).

LanthaScreen™ TR-FRET RAR beta Coactivator Assay Kit, goat

This kit contains Goat Tb-Anti-GST antibody; other kit components are the same as kit A15137:

The LanthaScreen® TR-FRET RAR (retinoic acid receptor) beta Coactivator Assay Kit provides a sensitive and robust method for highthroughput screening of potential RAR beta ligands as agonists of ligand-dependent coactivator recruitment. The kit uses a terbium (Tb)-labeled anti-GST antibody, a fluorescein-labeled coactivator peptide, and a human recombinant RAR beta ligand-binding domain (RAR beta-LBD) that is tagged with glutathione-S-transferase (GST) in a homogeneous, mix-and-read assay format.

When running the LanthaScreen® TR-FRET RAR beta coactivator assay in agonist mode to identify agonist compounds, RAR beta-LBD is added to ligand test compounds, followed by addition of a mixture of the fluorescein-coactivator peptide and Tb-labeled anti-GST antibody. After an incubation period at room temperature, the 520 nm/495 nm TR-FRET ratio is calculated and can be used to determine the EC50 from a dose response curve of the compound. Based on the biology of the RAR beta-coactivator peptide interaction, this ligand EC50 is a composite value representing the amount of ligand required to bind to receptor, effect a conformational change, and recruit coactivator peptide (Figure 1).

Contents and Storage:
The LanthaScreen® TR-FRET RAR beta Coactivator Assay Kit contains RAR beta-LBD (GST) protein, fluorescently labeled SRC2-2 coactivator peptide, Tb-labeled anti-GST antibody, and buffers. Store components as indicated in the assay protocol (-80°C, -20°C, or +4°C).