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)).
Elisa kits for mouse and human insulin
Manufactured by Mercodia
Mercodia's ELISA kits for mouse and human insulin are laboratory assays designed to quantify insulin levels in biological samples. These kits utilize the Enzyme-Linked Immunosorbent Assay (ELISA) technique to detect and measure insulin concentrations.
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2 protocols using elisa kits for mouse and human insulin
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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