Biochemical Nuclear Receptor Assays - Kits

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).

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 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

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 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.

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).

LanthaScreen™ TR-FRET Glucocorticoid Receptor Coactivator Assay Kit

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

Agonist Mode
When running the LanthaScreen TR-FRET Glucocorticoid Receptor Coactivator Assay in agonist mode (to identify agonist compounds), GR-LBD is added to ligand test compounds, followed by the addition of a mixture of fluorescein-coactivator peptide and Tb-labeled anti-GST antibody. After incubation at room temperature, the 520/495 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 GR-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).

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

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.

LanthaScreen™ TR-FRET PPAR alpha Competitive Binding Assay Kit, rabbit

This kit contains Rabbit Tb-Anti-GST antibody; other kit components are the same as kit PV4892:

The LanthaScreen® TR-FRET PPAR alpha Competitive Binding Assay provides a sensitive and robust method for high-throughput screening (HTS) of ligands for peroxisome proliferator-activated receptor alpha (PPAR alpha). The kit uses a terbium-labeled anti-GST antibody, a fluorescent small-molecule pan-PPAR ligand (Fluormone™ Pan-PPAR Green), and a human PPAR alpha ligand-binding domain (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 PPAR alpha Competitive Binding Assay, Fluormone™ Pan-PPAR Green is added to ligand test compounds followed by addition of a mixture of the PPAR alpha-LBD and terbium anti-GST antibody. When the Fluormone™ Pan-PPAR Green is bound to PPAR alpha, energy transfer from the terbium-labeled antibody to the tracer occurs, and a high TR-FRET ratio is observed. Competitive ligand binding to PPAR alpha is detected by a test compound’s ability to displace the tracer, which results in a loss of FRET between the antibody and the tracer. After an incubation period at room temperature, the 520 nm/495 nm TR-FRET ratio is calculated and can be used to determine the IC50 from a dose response curve of the compound (Figure 1). This type of binding assay is analogous to radioligandbased assays, except that it eliminates handling of radioactivity and enables a homogeneous "addition-only" format.

LanthaScreen™ TR-FRET Glucocorticoid Receptor Competitive Binding Kit

The LanthaScreen® TR-FRET Glucocorticoid Receptor Competitive Binding Assay Kit provides a sensitive and robust method for high-throughput screening (HTS) of ligands for glucocorticoid receptor (GR). The kit uses a human GR ligand-binding domain (GR-LBD) tagged with glutathione S-transferase (GST) (also available separately), a terbium-labeled anti-GST antibody, and a fluorescent small-molecule GR ligand (Fluormone™ GS1) in a homogeneous mix-and-read assay format.

The Lanthascreen Gluococorticoid Receptor 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 Androgen Receptor Coactivator Assay Kit

The LanthaScreen® TR-FRET Androgen Receptor Coactivator Assay Kit provides a sensitive and robust method for high-throughput screening of potential androgen receptor (AR) ligands as agonists of coactivator recruitment or antagonists of agonist-dependent coactivator recruitment. In both the agonist and antagonist modes, the homogeneous mix-and-read assay uses a rat AR ligand-binding domain (AR-LBD) tagged with hexahistidine and glutathione-S-transferase (GST) (also available separately), a terbium (Tb)-labeled anti-GST antibody, and a fluorescein-labeled coactivator peptide.

Agonist Mode
When the LanthaScreen TR-FRET Androgen Receptor Coactivator Assay is run in agonist mode (to identify agonist compounds), AR-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 nm emission ratio is calculated and used to determine the EC50 from a dose response curve of the compound. 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).

Antagonist Mode
When the LanthaScreen TR-FRET Androgen Receptor Coactivator Assay is run in antagonist mode (to identify antagonist compounds), AR-LBD is added to ligand test compounds, followed by addition of a mixture of DHT agonist, fluorescein-coactivator peptide, and Tb-anti-GST antibody. The concentration of DHT agonist used in this mode is the EC80 concentration as determined by first running the assay in agonist mode. An example of data produced in the antagonist mode is shown in the figure below.

PolarScreen™ ER Alpha Competitor Assay, Red

The PolarScreen™ ER (Estrogen Receptor) Alpha Competitor Assay Kit, Red, 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 EL 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.

PolarScreen™ ER Alpha Competitor Assay, Red, 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 EL 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 EL 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.

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 FXR Coactivator Assay Kit, goat

This kit contains Goat Tb-Anti-GST antibody; other kit components are the same as kit A15140:

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 PPAR gamma Competitive Binding Assay Kit, rabbit

This kit contains Rabbit Tb-Anti-GST antibody; other kit components are the same as kit PV4894:

The LanthaScreen® TR-FRET PPAR gamma Competitive Binding Assay provides a sensitive and robust method for high-throughput screening (HTS) of ligands for peroxisome proliferator-activated receptor-gamma (PPAR gamma). The kit uses a terbium-labeled anti-GST antibody, a fluorescent small-molecule pan-PPAR ligand (Fluormone™ Pan-PPAR Green), and human PPAR gamma ligand-binding domain (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 PPAR gamma Competitive Binding Assay, Fluormone™ Pan-PPAR Green is added to ligand test compounds followed by addition of a mixture of the PPAR gamma-LBD and terbium anti-GST antibody. When the Fluormone™ Pan-PPAR Green is bound to PPAR gamma, energy transfer from the terbium-labeled antibody to the tracer occurs, and a high TR-FRET ratio is observed. Competitive ligand binding to PPAR gamma is detected by a test compound’s ability to displace the tracer, which results in a loss of FRET between the antibody and the tracer. After an incubation period at room temperature, the 520 nm/495 nm TR-FRET ratio is calculated and can be used to determine the IC50 from a dose response curve of the compound (Figure 1). This type of binding assay is analogous to radioligand-based assays, except that it eliminates handling of radioactivity and enables a homogeneous, "addition-only" format.

LanthaScreen™ TR-FRET Progesterone Receptor Coactivator Assay Kit

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

Agonist Mode
When using the LanthaScreen TR-FRET Progesterone Receptor Coactivator Assay in agonist mode (to identify agonist compounds), PR-LBD is added to ligand test compounds, followed by the addition of a mixture of fluorescein-labeled coactivator peptide and Tb-labeled anti-GST antibody. After an incubation period at room temperature, the TR-FRET ratio of 520/495 nm is calculated and can be used to determine the EC50 from a dose response curve of the compound. Based on the biology of the PR-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).

Antagonist Mode
When using the LanthaScreen™ TR-FRET Progesterone Receptor Coactivator Assay in antagonist mode (to identify antagonist compounds), PR-LBD is added to ligand test compounds followed by addition of a mixture of agonist, fluorescein-labeled coactivator peptide, and terbium-labeled anti-GST antibody (see figure). The concentration of agonist used in this mode is the EC80 concentration as determined by first running the assay in agonist mode.