Mouse rat igf 1 igf 1 quantikine elisa kit

The postnatal leptin surge in our FVB mice was initially characterized with serum collected from pups aged postnatal day (PND) 1, 5, 8, 11, and 16. Ahima et al. (17 (link)) and results from a pilot study dictated our selection of PND 1, 5, 8, 11, and 16 for our CR20 study (see Methods 2.3). Whole blood was incubated on ice for 1 hour and centrifuged at 3200 x g for 20 minutes at 4°C. Serum was collected on ice and stored at -20°C until use. Serum leptin levels were measured at a 1:10 dilution using the Mouse/Rat Leptin Quantikine ELISA Kit (RNDSystems, MOB00). Serum IGF-1 was measured at a 1:500 dilution using R&D Systems Mouse/Rat IGF-I/IGF-1 Quantikine ELISA Kit (MG100). Serum pituitary hormones, LH, PRL, ACTH, FSH, and TSH, were measured with at 1:2.5 dilution with Milliplex’s MAP Mouse Pituitary Magnetic Bead Panel - Endocrine Multiplex Assay (MPTMAG-49K). Serum adipokines (IL-6, Insulin, MCP-1, PAI-1, Resistin, TNF-α) were measured undiluted with Milliplex’s MAP Mouse Adipokine Magnetic Bead Panel (MADKMAG-71K). Serum GH was measured at 1:2.5 dilution with Milliplex’s MAP Rat Pituitary Magnetic Bead Panel (RPTMAG-86K-01).

Plasma samples were collected into heparin-coated tubes at six time points (ZT 1, ZT 5, ZT 9, ZT 13, ZT 17, ZT 21) throughout a 24-h period. Plasma leptin concentrations were determined by ELISA (mouse leptin ELISA kit, 90030, Crystal Chem, IL). Plasma insulin concentrations were determined by ELISA (Ultra Sensitive Mouse Insulin ELISA Kit, 90080, Crystal Chem, IL). Plasma IGF-1 concentrations were determined by ELISA (Mouse/Rat IGF-I/IGF-1 Quantikine ELISA Kit, MG100, R&D Systems, MN). Plasma corticosterone concentrations were determined by ELISA (corticosterone ELISA Kit, K014-H1, Arbor Assays, MI). Plasma sodium, potassium, and chloride levels were measured using the Abbott point of care i-STAT Alinity system.

MRM were isolated and cultured from RA and OIR mice at P17. The secretion of IGF1 from cultured retinal microglia was detected using Mouse/Rat IGF-I/IGF-1 Quantikine ELISA Kit (R&D systems, MG100) according to the manufacturer’s instructions.