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Was measured using Insulin (Mouse) Ultrasensitive ELISA kit (ALPCO Diagnostics, Salem, NH) according to manufacture's protocol.
Saturation of IR and IGF-1R was measured using Insulin signaling phospho/total assay.
The supernatants were collected, and secreted insulin was measured using insulin ELISA kits (for mouse insulin from Millipore and for human insulin from LifeTech).
Insulin in mouse plasma, in islet culture samples and in pancreatic extracts was measured using insulin enzyme-linked immunosorbent assay (ELISA, Crystal Chem, Downers Grove, IL, USA).
Insulin concentration in blood sera was measured using Insulin (Mouse) Ultrasensitive ELISA kit (ALPCO Diagnostics, Salem, NH, USA) according to manufacture's protocol.
Insulin, IGF1, leptin, and triglyceride concentration in blood plasma, were measured using Insulin (Mouse) Ultrasensitive ELISA kit (ALPCO Diagnostics, Salem, NH, USA), IGF1 (Mouse/Rat) ELISA kit (ALPCO), Mouse Leptin ELISA kit (Crystal Chem Inc ,Downers Grove, IL, USA), and Triglyceride Colorimetric Assay kit (Cayman Chemical Company, Ann Arbor, MI, USA), respectively.
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Serum insulin was measured using insulin-specific radioimmunoassay kits for rats (Linco Research).
Insulin was measured using mouse insulin ELISA (Ultrasensitive Mouse Insulin ELISA, Mercodia, Sweden).
Insulin sensitivity was measured using the insulin sensitivity index, which is 22.5/(fasting insulin × [fasting glucose/18.0]), the reciprocal of which is the homeostasis model assessment of insulin resistance (HOMA-IR) (20).
Plasma insulin was measured using ultrasensitive insulin enzyme-linked assay (Mercodia, Uppsala, Sweden).
Our results were consistent with those of large-scale studies that found elevated serum calcium levels to be associated with insulin resistance, measured using fasting insulin or homeostatic model assessment [2], [27], [28].
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