规格
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Assay Entry:
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Biochemical competitive binding |
Product Line:
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LanthaScreen® |
Readout:
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End Point |
Shipping Condition:
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Dry Ice |
Target Entry:
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PPARA, PPAR alpha, NR1C1 |
Technique:
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TR-FRET |
Validated Application:
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Competitive Binding Assay |
Label or Dye:
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Tb (Terbium) |
Detection Method:
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Fluorescent |
For Use With (Equipment):
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Microplate Reader |
Format:
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384-well plate |
储存
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The LanthaScreen® TR-FRET PPAR alpha Competitive Binding Assay Kit contains PPAR alpha-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). |
This kit contains Goat Tb-Anti-GST antibody; other kit components are the same as kit A15143:
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.