Product Overview :
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.
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.
Applications :
• Monitor glucocorticoid signaling pathway activity. • Screen activators or inhibitors of the glucocorticoid signaling pathway.
Storage :
>1 year from date of receipt when stored at -20°C
Kit Components :
Reporter (Component A): GAL4/UAS luciferase reporter vector + constitutively expressing Renilla luciferase vector; 500 µl (60 ng DNA/µl); -20°C Negative Control Reporter (Component B): Non-inducible luciferase vector + constitutively expressing Renilla luciferase vector; 500 µl (60 ng DNA/µl); -20°C GAL4 DBD-GR (Component C): Expression vector for ligand binding domain of the glucocorticoid receptor + GAL4 DNA binding domain; 250 µl (100 ng DNA/µl); -20°C Negative Control Expression vector (Component D): Expression vector with GAL4 DNA binding domain only; 250 µl (100 ng DNA/µl); -20°C
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