All metabolic tests were performed after 6 h of fasting [32] . For insulin clearance during an oral glucose challenge, fasting blood glucose and blood samples (50 µL) were collected from the tail vein after 6 hours of fasting. Mice were then given a 4 g/kg glucose dose by oral gavage and subsequent blood samples (50 µL) were collected from the tail vein at 10, 60, and 120 minutes post-gavage. For insulin clearance during an insulin challenge, mice were given human insulin (1 U/kg, NovoRapid) or human C-peptide (50 µg/kg, Sigma) by intraperitoneal injection and blood samples were collected by tail vein sampling at 0, 5, 30, and 60 minutes post-injection. All blood samples were kept on ice after collection and then centrifuged at 10,000 g for 10 min at 4˚C. Plasma was collected into fresh tubes and stored at -80˚C. Mouse insulin and C-peptide were detected by multiplex ELISA (Millipore) kit in the plasma samples collected during the oral glucose challenge. human insulin (Mercodia) and human C-peptide (Millipore) were detected by ELISA kits in the plasma samples collected during the human insulin or human C-peptide challenges, respectively.
Human c peptide
Manufactured by Merck Group
The Human C-peptide is a laboratory assay product used to measure the level of C-peptide in human samples. C-peptide is a byproduct of insulin production and its measurement can provide insights into insulin secretion and diabetes management.
Automatically generated - may contain errors
2 protocols using human c peptide
1
Glucose and Insulin Clearance Assays
2
Diabetic Mice C-peptide Supplementation
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Four weeks after streptozotocin injection, a group of diabetic mice (DM+CP) were anesthetized with an intraperitoneal injection of 2.5% Tribromoethanol (240 mg/kg, Sigma-Aldrich) and supplemented with human C-peptide (American Peptide Company, Inc, Sunnyvale, CA, USA) using an Alzet Mini-Osmotic Pump 2004 (DURECT, Cupertino, CA, USA) with a delivery rate of 35 pmol/min/kg. Anesthesia was monitored via the pain reflex reaction (lack of response to hind limb toe pinch). A second group of diabetic mice (DM+Ins; HGM) was supplied with human recombinant insulin (Sigma-Aldrich), with a delivery rate of 58.4 pmol/min/kg to mimic hyperglycemic memory. To assess the effect of C-peptide treatment on HGM, a third group of diabetic mice (DM+Ins+CP; HGM+CP) was supplemented with a mixture of C-peptide and insulin, also via the osmotic pump. Diabetic (DM) and control mice groups also underwent sham operations. The lack of mouse C-peptide in appropriate groups was measured using a C-peptide Enzyme Immunoassay Kit (RayBiotech, Norcross, GA, USA). Supplemented human C-peptide and/or insulin levels in serum were measured using ELISA kits specific for each (Millipore). The vital statistics of each mouse were recorded throughout the experiment.
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