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Thermo Scientific™ LanthaScreen™ TR-FRET PPAR gamma Competitive Binding Assay Kit, rabbit

Catalog No. A15145
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400 x 40 μL assays
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Description

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).

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.

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Specifications

Color Green
Conjugate Tb (Terbium)
Detection Method Fluorescence
For Use With (Equipment) Microplate Reader
Quantity 400 x 40 μL assays
Packaging 384-well Plate
Content And Storage • Tb-anti-GST Antibody (Rabbit) (store at -20°C)
• PPAR gamma-LBD, GST (-80°C)
• Fluormone Pan-PPAR Green (-20°C)
• TR-FRET PPAR Assay Buffer (4°C)
• 1M DTT (-20°C or -80°C)
Assay Category Biochemical Assay
Druggable Target Nuclear Receptors
Readout TR-FRET (Binding)
Product Line LanthaScreen
For Use With (Application) Nuclear Receptor Assay, TR-FRET
Ligand Pan-PPAR
Product Type TR-FRET PPAR Gamma Competitive Binding Assay Kit
No. of Assays 400 x 40 μL Assays
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For Research Use Only. Not for use in diagnostic procedures.

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