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Invitrogen™ LanthaScreen™ TR-FRET PPAR gamma Competitive Binding Assay Kit, goat
Description
This kit contains Goat Tb-Anti-GST antibody; other kit components are the same as kit A15145:
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.
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.
Specifications
Specifications
| Conjugate | Tb (Terbium) |
| Detection Method | Fluorescence |
| For Use With (Equipment) | Microplate Reader |
| Quantity | 400 x 40 μL |
| Packaging | 384-well Plate |
| Content And Storage | The LanthaScreen™ TR-FRET PPAR gamma Competitive Binding Assay Kit contains PPAR gamma-LBD (GST) protein, Fluormone™ Pan-PPAR Green, terbium-labeled anti-GST antibody, and buffers. Store components as indicated in the assay protocol (-80°C, -20°C, or +4°C). |
| Assay Entry | Biochemical competitive binding |
| Readout | End Point |
| Shipping Condition | Dry Ice |
| Target Entry | PPARG, PPAR gamma, NR1C3 |
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Safety and Handling
WARNING: Cancer - www.P65Warnings.ca.gov
For Research Use Only. Not for use in diagnostic procedures.
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