规格
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Assay Entry:
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Biochemical coregulator interaction |
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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RAR beta, NR1B2, RARB |
Technique:
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TR-FRET |
Validated Application:
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Co-Factor Interaction Assay |
Label or Dye:
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Tb (Terbium), FITC (Fluorescein) |
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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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 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).