规格
|
Assay Entry:
|
Biochemical coregulator interaction |
Gene ID (Entrez):
|
5467 |
Label or Dye:
|
Tb (Terbium) |
Product Line:
|
LanthaScreen® |
Readout:
|
End Point |
Shipping Condition:
|
Dry Ice |
Target Entry:
|
PPAR delta, NR1C2, PPARD |
Technique:
|
TR-FRET |
Validated Application:
|
Co-Factor Interaction Assay |
Detection Method:
|
Fluorescent |
For Use With (Equipment):
|
Microplate Reader |
Format:
|
384-well plate |
储存
|
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 A15134:
The LanthaScreen® TR-FRET PPAR (peroxisome proliferator activated receptor) delta Coactivator Assay Kit provides a sensitive and robust method for high-throughput screening of potential PPAR delta 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 PPAR delta ligand-binding domain (PPAR delta-LBD) that is tagged with glutathione-S-transferase (GST), in a homogeneous mix-and-read assay format.
When running the LanthaScreen® TR-FRET PPAR delta Coactivator Assay in agonist mode (to identify agonist compounds), PPAR delta-LBD is added to ligand test compounds followed by addition of a mixture of the fluorescein-labeled coactivator peptide and Tb-labeled anti-GST antibody. After an incubation period at room temperature, the TR-FRET ratio of 520/495 is calculated and can be used to determine the EC50 from a dose response curve of the compound. Based on the biology of the PPAR delta-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 (see Figure 1).
Contents and Storage:
The LanthaScreen® TR-FRET PPAR delta Coactivator Assay Kit contains PPAR delta-LBD (GST) protein, fluorescently labeled C33 coactivator peptide, Tb-anti-GST antibody, and buffers. Store components as indicated in the assay protocol (-80°C, -20°C or 4°C).