Glucocorticoids are a family of steroids necessary for the regulation of energy metabolism and the immune and inflammatory responses. These compounds exert their effect through their interaction with the Glucocorticoid Receptor (GR) and that complex's subsequent association with DNA. All normal mammalian tissues examined to date have been shown to contain GR. The human GR exists in two forms, alpha and beta, which are thought to be the result of alternative splicing of a single gene. Sequence analysis indicates that alpha and beta forms of human GR are 777 and 742 amino acids long, respectively. They are identical up to residue 727, after which they diverge. After ligand binding, the 94kD GR alpha isoform translocates from the cytoplasm to the nucleus where it regulates gene expression. In contrast, the 90kD GR beta isoform does not appear to bind either glucocorticoid agonists or antagonists, and has been localized predominantly in the nucleus independent of hormone treatment in some human cell lines. Studies suggest that human GR alpha has a greater affinity for GR response elements (GREs) than GR beta only when in the ligand bound state.
Applications:
Suitable for use in ELISA, Western Blot and Immunoprecipitation. Other applications not tested.
Recommended Dilution:
Western Blot: 4ug/ml. Detects ~90kD protein representing GR in HeLa cell lysate.
Optimal dilutions to be determined by the researcher.
Storage and Stability:
Lyophilized powder may be stored at -20 degrees C for short-term only. Reconstitute with sterile 40-50% glycerol, PBS, aliquot and store at -20 degrees C. Reconstituted product is stable for 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
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