规格
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Assay Entry:
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Biochemical coregulator interaction |
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Label or Dye:
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Tb (Terbium) |
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Product Line:
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LanthaScreen® |
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Readout:
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End Point |
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Shipping Condition:
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Dry Ice |
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Target Entry:
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THRA, TR alpha, NR1A1 |
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Technique:
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TR-FRET |
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Validated Application:
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Co-Factor Interaction Assay |
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Detection Method:
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Fluorescent |
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For Use With (Equipment):
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Microplate Reader |
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Format:
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384-well plate |
储存
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Store in ultra-cold freezer (-68 to -85°C). |
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).
