Primary monoclonal antibodies against GRα, GRβ, phospho-p38MAPK, phospho-JNK and β-actin, as well as a horseradish peroxidase-labeled secondary goat anti-mouse IgG antibody, were purchased from Santa Cruz Biotechnology (USA). IL-1β was purchased from PeproTech (England) and RPMI1640 culture medium was purchased from GIBCO (USA). The specific inhibitors of the p38 MAPK pathway (SB203580), ERK pathway (PD98059), PI3K pathway (LY294002), PKC pathway (Ro31-8220) and JNK pathway (SP600125) were all purchased from Calbiochem (USA). [3H]-Dexamethasone was obtained from Amersham (USA). Dexamethasone was purchased from Sigma (USA). The primers and Taqman probes targeting GRα, GRβ, p38 MAPK and JNK were designed and synthesized by Applied Biosystems (USA). All other chemicals used in the study were of analytical grade and obtained from commercial sources.
3h dexamethasone
Manufactured by Cytiva
Sourced in United States, United Kingdom
[3H]-Dexamethasone is a radiolabeled synthetic glucocorticoid used as a research tool in various experimental settings. It binds to glucocorticoid receptors and can be used to study the receptor's distribution, kinetics, and function.
Automatically generated - may contain errors
Lab products found in correlation
Horseradish peroxidase labeled secondary goat anti mouse igg antibody, by Santa Cruz Biotechnology (1 mentions)
Rpmi 1640 culture medium, by Thermo Fisher Scientific (1 mentions)
Primary monoclonal antibody, by Santa Cruz Biotechnology (1 mentions)
Il 1β, by Thermo Fisher Scientific (1 mentions)
Dexamethasone (dex), by Merck Group (1 mentions)
2 protocols using 3h dexamethasone
1
Glucocorticoid Receptor Pathway Investigation
2
Dexamethasone Receptor Binding Assay
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Cell suspensions (1×10 6 cells) were incubated (120 min, 37°C) with [ 3 H]dexamethasone (Amersham, UK; specific activity 40 Ci/mmol) at five different concentrations ranging from 7.5-120 nmol/l. Triplicate incubations were performed in the absence and in the presence of 100-fold molar excess of unlabeled dexamethasone (Sigma-Aldrich, Germany) to determine total and nonspecific binding, respectively, and to calculate specific binding from their difference. The maximal number of receptor sites per cell (Bmax) and equilibrium dissociation constant (KD) were calculated by computer-assisted nonlinear regression. Hormone-binding potency of the receptor was calculated as Bmax/KD ratio.
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