Bio-Rad protein assay kit was purchased from Bio-Rad; sodium pyruvate, malic acid, succinic acid, ascorbic acid, N,N,N′,N′-tetramethyl-p-phenylenediamine (TMPD), palmitoyl-L-carnitine, rotenone, antimycin A, oligomycin, coenzyme A trilithium salt (CoA-SH), acetyl-CoA, oxaloacetic acid, thiamine pyrophosphate (TPP), 5,5-dithiobis-(2-nitrobenzoic acid) (DTNB), 2,6-dichlorophenolindophenol (DCPIP), carbonyl cyanide-4-(trifluoromethoxy)phenylhydrazone (CCCP), phosphoenolpyruvate (PEP), ADP, ATP, NAD+, NADH, NADPH, KCN, pyruvate kinase, lactate dehydrogenase, cytochrome c, and decylubiquinone were from Sigma; KO was a generous gift of Aker BioMarine ASA (Oslo, Norway). Antibodies against AAC and UCP2 were from Santa Cruz Biotechnology (sc-11433 and sc-6526); antibodies against OXPHOS proteins were from Mitosciences (ab110413). Kits for the assay of triglycerides and total cholesterol were purchased from Futura System. Plasma insulin concentration was analyzed with a Mercodia Ultrasensitive Mouse Insulin kit. Luciferase ATP assay kit was from Sigma and Lipid Hydroperoxide (LPO) assay kit was from Merck. All other reagents were of analytical grade.
Ultrasensitive mouse insulin kit
Manufactured by Mercodia
Sourced in Sweden
The Ultrasensitive Mouse Insulin Kit is a laboratory assay used for the quantitative determination of insulin levels in mouse samples. The kit utilizes an enzyme-linked immunosorbent assay (ELISA) technique to measure insulin concentrations.
Automatically generated - may contain errors
Lab products found in correlation
Rotenone, by Merck Group (1 mentions)
Oxaloacetic acid, by Merck Group (1 mentions)
Thiamine pyrophosphate, by Merck Group (1 mentions)
Adenosine diphosphate (adp), by Merck Group (1 mentions)
Pyruvate kinase, by Merck Group (1 mentions)
Lactate dehydrogenase, by Merck Group (1 mentions)
Acetyl coa, by Merck Group (1 mentions)
Protein assay kit, by Bio-Rad (1 mentions)
Adenosine triphosphate (atp), by Merck Group (1 mentions)
Cytochrome c, by Merck Group (1 mentions)
Oligomycin, by Merck Group (1 mentions)
Antimycin a, by Merck Group (1 mentions)
Nadph, by Merck Group (1 mentions)
Ascorbic acid, by Bio-Rad (1 mentions)
Decylubiquinone, by Merck Group (1 mentions)
Palmitoyl l carnitine, by Merck Group (1 mentions)
Accu check, by Roche (1 mentions)
5 protocols using ultrasensitive mouse insulin kit
1
Biochemical Assay Protocol Compendium
2
Oral Glucose Tolerance Test in Mice
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For the OGTT, mice were fasted overnight (16 h), before starvation the total body weight, fat, and lean mass was recorded as well as free and total water contribution using the EchoMRI™ scan. Then, after 16 h the animals were again scanned using the EchoMRI™ scan and basal blood glucose was measured by Accu-check stripes and the OneTouch Vita blood glucose meter. Mice were given glucose bolus (1 g/kg body weight) by oral gavage. Blood glucose levels were measured using a OneTouch Vita blood glucose meter at −5 (to avoid stress bias), 0, 15, 30, 45, and 60 min after glucose. Blood drawn using heparinized capillary tubes was centrifuged at 2000 × g for 15 min at 4 °C. Plasma obtained was used for insulin level measurement by ELISA (Mercodia Ultrasensitive Mouse Insulin Kit).
3
Liraglutide Effects on Glucose and Insulin in Mice
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During the whole experiment, the body weight and blood glucose of mice were measured 5 times, before and 3, 7, 11, and 14 days after liraglutide administration. Glucose measurement was performed by a blood glucose meter (Accu‐Check, Roche) using blood collected from the tail vein of mice. At the end of the experiment, blood collected from the inferior vena cava after overnight fasting was centrifuged at 2000g for 15 minutes at 4 °C. Plasma obtained was used for insulin level measurement by ELISA (Ultrasensitive Mouse Insulin Kit; Mercodia, Uppsala, Sweden) according to the manufacturer's instructions.
4
Oral Glucose Tolerance Test in Mice
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For the oral glucose tolerance test (OGTT), mice were fasted overnight (16 hr), after which basal blood glucose was measured. Mice were given glucose (1 g/kg) by oral gavage. Blood glucose levels were measured using a OneTouch Vita blood glucose meter at 0, 15, 30, 45, and 60 min after glucose. Blood drawn using heparinized capillary tubes was centrifuged at 2000 g for 15 min at 4°C. Plasma obtained was used for insulin level measurement by ELISA (Mercodia Ultrasensitive Mouse Insulin Kit).
5
Oral Glucose Tolerance Test in Mice
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On the day of the experiment, food was removed in the morning at 7:00 (winter) or 8:00 (summertime). The OGTT was conducted after a fasting period of 6 h (i.e., at 13:00 or 14:00). Body weight was measured before the start of the experiment. Access to an RW was maintained during the OGTT time frame. In the RW groups, the last bout of intense exercise occurred 6-7 h before the ingestion of the glucose bolus, according to data on average RW revolutions at different ages (Supplementary Fig. 1). At time point zero, a glucose bolus of 1 g • kg À1 (5.5 • 10 3 mmol • kg À1 ) was administered orally. It consisted of 0.7 g • kg À1 (3.9 • 10 3 mmol • kg À1 ) unlabeled glucose and 0.3 g • kg À1 (1.6 • 10 3 mmol • kg À1 ) [6,6-2 H 2 ]glucose (a tracer). Total blood glucose (Lifescan Euroflash; Lifescan Benelux, Beerse, Belgium) was measured from the tail vein, from which blood spots were also collected. Insulin was measured according to manufacturer instructions (Ultrasensitive Mouse Insulin kit; Mercodia, Uppsala, Sweden). The fractional contribution of the administered tracer in blood glucose (described below) was measured in air-dried blood spots. Glucose and tracer enrichment were measured at 0, 15, 30, 45, 60, 75, 90, 105, and 120 min and insulin was measured at 0, 30, and 60 min.
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