GAL4 Reporter Kit (Glucocorticoid Receptor Pathway)

Catalog #
60522
$565 *
Size: 500 reactions
Qty
*US Pricing only. For international pricing, please contact your local distributor.
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Description

The Glucocorticoid Receptor Pathway Reporter Kit is designed for monitoring the activity of the glucocorticoid signaling pathway in cultured cells. The kit contains a transfection-ready expression vector for the glucocorticoid receptor ligand binding domain that is fused to the DNA binding domain (DBD) of GAL4 (GAL4 DBD-GR). This fusion construct activates firefly luciferase expression under the control of a multimerized GAL4 upstream activation sequence (UAS). This allows for specific detection of glucocorticoid-induced activation of the glucocorticoid receptor without the need for individual transcriptional targets and with low crossreactivity for other nuclear receptor pathways. The GAL4/UAS reporter is premixed with constitutively expressing Renilla (sea pansy) luciferase vector, which serves as an internal positive control for transfection efficiency.

The kit also includes a non-inducible firefly luciferase vector premixed with constitutivelyexpressing Renilla luciferase vector as a negative control. The non-inducible luciferase vector contains a firefly luciferase gene under the control of a minimal promoter, but without any additional response elements. The negative control is critical for determining pathway-specific effects and background luciferase activity.

Purchase of this cell line is for research purposes only; commercial use requires a separate license. View the full terms and conditions.

Synonyms
Glucocorticoid Receptor, GR, Nuclear receptor subfamily 3 group C member 1, NR3C1, GAL4, GAL4/UAS, Galectin, Cell based assay kit, cell-based assay, reporter assay kit
Product Info
Storage and Usage
Citations
Materials Required But Not Supplied
  •  Mammalian cell line and appropriate cell culture medium
  • 96-well tissue culture plate or 96-well tissue culture treated white clear-bottom assay 
    plate (Corning # 3610)
  • Transfection reagent for mammalian cell line. [We use Lipofectamine™ 2000 (Life 
    Technologies #11668027), however, other transfection reagents should work equally 
    well.]
  • Opti-MEM I Reduced Serum Medium (Life Technologies #31985-062)
  • TWO-step luciferase assay system:
  • TWO-Step Luciferase (Firefly & Renilla) Assay (BPS Bioscience #60683): This system 
    assays cells directly in growth medium. It can be used with any luminometer. Automated 
    injectors are not required.
  • Luminometer
Format
COMPONENTS:

Background
The glucocorticoid signaling pathway plays an important role in development, fluid homeostasis, cognition, immune response and metabolism. Glucocorticoids are a class of steroid hormones that bind to the glucocorticoid receptor, causing it to translocate to the nucleus. Upon translocation, the receptor can regulate the transcription of a large number of genes, including those that regulate glucose metabolism and inflammatory responses.
References
Paguio A, et al. (2010) Improved Dual-Luciferase Reporter Assays for Nuclear Receptors. Curr Chem Genomics. 4: 43-49.