Insulin, hRANKL, and hOPG from serum or culture media were assayed using ELISA kits for mouse and human insulin (Mercodia Inc., Winston Salem, NC) (12 (link), 87 (link), 91 (link), 92 (link)), human RANKL (abcam, Waltham, MA), and human OPG (abcam), respectively. IL-1β and IL-6 production in monocytes was determined using ELISA, per the manufacturer’s instructions (BioLegend, San Diego, CA). For Western blot analysis, protein extracts (20 to 40 μg) from human and mouse islets treated with or without cytokines in the presence or absence of IgG, OPG (100 ng/ml), or DMB (100 ng/ml) for 24 hours were analyzed by immunoblotting using antibodies against p-NF-κB-Ser536, p-STAT1-Ser727 (Cell Signaling Technology, Danvers, MA), tubulin (EMD Millipore-Calbiochem, Burlington, MA), and glyceraldehyde-3-phosphate dehydrogenase (Sigma-Aldrich, St. Louis, MO) (table S4). Quantitative densitometry of digitalized blots was performed using the ImageJ program (12 (link), 87 (link), 88 (link), 90 (link)–92 (link)).
Human rankl
Manufactured by Abcam
Human RANKL is a recombinant protein that corresponds to the extracellular domain of the human receptor activator of nuclear factor-kappa B ligand (RANKL). RANKL is a member of the tumor necrosis factor (TNF) superfamily and plays a crucial role in the regulation of bone metabolism, immune function, and other physiological processes.
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Lab products found in correlation
2 protocols using human rankl
1
Quantifying Insulin, RANKL, and OPG Levels
2
Quantifying Insulin, RANKL, and OPG Levels
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Insulin, hRANKL, and hOPG from serum or culture media were assayed using ELISA kits for mouse and human insulin (Mercodia Inc., Winston Salem, NC) (12 (link), 87 (link), 91 (link), 92 (link)), human RANKL (abcam, Waltham, MA), and human OPG (abcam), respectively. IL-1β and IL-6 production in monocytes was determined using ELISA, per the manufacturer’s instructions (BioLegend, San Diego, CA). For Western blot analysis, protein extracts (20 to 40 μg) from human and mouse islets treated with or without cytokines in the presence or absence of IgG, OPG (100 ng/ml), or DMB (100 ng/ml) for 24 hours were analyzed by immunoblotting using antibodies against p-NF-κB-Ser536, p-STAT1-Ser727 (Cell Signaling Technology, Danvers, MA), tubulin (EMD Millipore-Calbiochem, Burlington, MA), and glyceraldehyde-3-phosphate dehydrogenase (Sigma-Aldrich, St. Louis, MO) (table S4). Quantitative densitometry of digitalized blots was performed using the ImageJ program (12 (link), 87 (link), 88 (link), 90 (link)–92 (link)).
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