At 12 weeks post-surgery all mice were overnight fasted and euthanized by decapitation. 500 µl of trunk blood was collected in microtubes containing 83.5 µl EDTA (Sigma, St. Louis, MO) and 15 µl of a Protease inhibitor cocktail (5 µl of each of the following: Protease inhibitor, Sigma, St. Louis, MO; DPP-IV inhibitor, EMD Millipore, St. Charles, MO; Pefabloc SC, Roche, Indianapolis, IN) and immediately centrifuged at 4° C and 3000 RPM for 10 minutes to separate the plasma from the whole blood. Plasma aliquots were frozen in liquid nitrogen, and stored at −80° C prior to processing. Plasma was subjected to ELISA for measurement of insulin and leptin concentrations (MMHMAG-44K Milliplex map mouse metabolic hormone magnetic bead panel – metabolism multiplex assay, EMD Millipore, St. Charles, MO). One drop of unprocessed tail vein blood was used for baseline glucose measurement (Onetouch Ultra Glucometer, LifeScan INC, Milpitas, CA; Onetouch Ultra Strips, LifeScan INC, Milpitas, CA). One drop of unprocessed trunk blood was used for baseline glucose measurement (Onetouch Ultra Glucometer, LifeScan INC, Milpitas, CA; Onetouch Ultra Strips, LifeScan INC, Milpitas, CA).
Onetouch ultra strips
Manufactured by LifeScan
Sourced in United States
Onetouch Ultra Strips are test strips designed for use with Onetouch Ultra blood glucose monitoring systems. They are used to measure blood glucose levels.
Automatically generated - may contain errors
Lab products found in correlation
Onetouch ultra glucometer, by LifeScan (6 mentions)
Mmhmag 44k milliplex map mouse metabolic hormone magnetic bead panel metabolism multiplex assay, by Merck Group (3 mentions)
Pefabloc sc, by Roche (3 mentions)
Protease inhibitor, by Merck Group (3 mentions)
Novolin r, by Novo Nordisk (3 mentions)
Dpp4 inhibitor, by Merck Group (2 mentions)
Ethylene diamine tetraacetic acid (edta), by Merck Group (2 mentions)
Ddp 4 inhibitor, by Merck Group (1 mentions)
Mouse insulin elisa kit, by Crystal Chem (1 mentions)
Onetouch ultra2, by LifeScan (1 mentions)
9 protocols using onetouch ultra strips
1
Plasma Metabolic Hormone Analysis in Mice
2
Intraperitoneal Glucose Tolerance Test
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Six weeks after surgery, intraperitoneal glucose tolerance was tested in overnight (15–17 h) food deprived mice by administering 2 g/kg of α-d -glucose (10% in sterile water, i.p.). Tail blood was analyzed by a glucometer (Onetouch Ultra Glucometer, LifeScan INC, Milpitas, CA; Onetouch Ultra Strips, LifeScan INC, Milpitas, CA). At 5 min before the injection, larger samples of 100 μl of whole blood were collected using heparinized capillary tubes (Fisherbrand Microhematocrit Capillary Tubes, Thermo Fisher Scientific, Waltham, MA) into centrifuge tubes containing 4.5 μl of a Protease inhibitor cocktail (1.5 μl of each of the following: Protease inhibitor, Sigma, St. Louis, MO; DDP-IV inhibitor, EMD Millipore, St. Charles, MO; Pefabloc SC, Roche, Indianapolis, IN) and immediately centrifuged at 4 °C and 3000 RPM for 10 min to separate the plasma from the whole blood. Plasma aliquots were frozen in liquid nitrogen and stored at −80 °C prior to processing. Plasma was subjected to ELISA for measurement of insulin and leptin concentrations (MMHMAG-44K Milliplex map mouse metabolic hormone magnetic bead panel – metabolism multiplex assay, EMD Millipore, St. Charles, MO).
3
CD3 Antibody Treatment Modulates Metabolic Profile
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Animals were purchased and maintained under specific pathogen‐free conditions at the central animal facility of the Hannover Medical School (Hannover, Germany). BALB/c Rag1−/− mice were bred in‐house. We fed 6‐8‐week‐old mice a normal chow diet (NCD) from ssniff (Altromin standard diet #1324 total pathogen free; Altromin) or an HF‐HC diet (ssniff EF R/M D12330 mod. */Surwit + 1% cholesterol; ssniff) with 45 g/L 55% fructose/45% sucrose (Sigma) in the drinking water for 16 weeks. Fifty micrograms of a non‐Fc receptor binding anti‐CD3 monoclonal antibody (anti‐CD3 F[ab])2’) (145‐2C11; Bio‐X‐Cell) or isotype control (Bio‐X‐Cell) was intraperitoneally injected into mice for 5 consecutive days during week 16 of diet feeding. For the glucose tolerance test, 16‐hour‐fasted mice received 1 mg glucose/g body weight. Blood glucose level was measured in blood collected from the mouse tail vein with OneTouchUltra strips (LifeScan).
4
Glucose Tolerance and GLP-1 Assay in Mice
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Male C57BL/6 (8 weeks old) were fasted for 16 h before glucose loading. Dapagliflozin (1 mg/kg) or HM41322 (0.3, 1, or 3 mg/kg) was orally administered to the mice 30 min before glucose loading (2 g/kg, 10 ml/kg). Blood glucose levels were determined using OneTouch® Ultra strips (Life Scan, Inc., USA) and tail-nick blood collected during, prior to, and 0, 15, 30, 60, and 120 min after glucose loading. Glucose AUC0–120min was calculated via the oral glucose tolerance test (OGTT) using the trapezoidal rule. Inhibiting SGLT1 improves glucose control by reducing intestinal glucose absorption and stimulating glucagon-like peptide-1 (GLP-1) release [19 (link)]. Thus, for determining total GLP-1, plasma was collected via the subclavian vein at 30 min after glucose loading. GLP-1 was analyzed using an ELISA kit for multispecies (Millipore, Cat no. EZGLP1T-36K).
5
Metabolic Profiling of Mice on High-Fat Diet
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Glucose tolerance tests (GTT) were performed a few weeks after the first period in the metabolic chambers (time after exposure to high-fat diet: 93 ± 4 days, referred to as 13 weeks, in females; 89 ± 6 days, referred to as 13 weeks, in males). Each GTT took place between 09:00–12:00 h after 3–5 hours of food deprivation. Mice were given an i.p. injection of α-D-glucose at a dose of 1.5 g/kg body weight, and blood glucose was measured before and at 15, 30, 60, and 120 minutes after injection by taking a small sample of blood from the tail vein and assessing it with glucose strips and a glucometer (Onetouch Ultra Strips and Onetouch Ultra Glucometer, LifeScan INC, Milpitas, CA), which can read blood glucose concentrations up to 600 mg/dL.
Insulin tolerance tests (ITT) were performed just before the second period in the metabolic chambers (time after exposure to high-fat diet: 201 ± 1 days, referred to as 29 weeks, in females; 174 ± 27 days, referred to as 25 weeks, in males). Each ITT took place between 09:00–12:00 h after 3–5 hours of food deprivation. Mice were given an i.p. injection of insulin (Novolin R, Novo Nordisk Inc., Plainsboro, NJ) at a dose of 0.6U/kg body weight, and blood glucose was measured with the same methods used for the GTT.
Insulin tolerance tests (ITT) were performed just before the second period in the metabolic chambers (time after exposure to high-fat diet: 201 ± 1 days, referred to as 29 weeks, in females; 174 ± 27 days, referred to as 25 weeks, in males). Each ITT took place between 09:00–12:00 h after 3–5 hours of food deprivation. Mice were given an i.p. injection of insulin (Novolin R, Novo Nordisk Inc., Plainsboro, NJ) at a dose of 0.6U/kg body weight, and blood glucose was measured with the same methods used for the GTT.
6
Plasma Metabolic Hormone Analysis in Fasted Mice
7
Fasting Blood Glucose and Insulin Resistance in Mice
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After 6 h of fasting, mouse blood was collected via tail nick to measure FBG using a OneTouch Ultra 2 glucometer with OneTouch Ultra strips (LifeScan Inc.). Submandibular blood was collected using a heparinized capillary (Kimble) and plasma was isolated by centrifugation ( at 4°C) and stored at until analysis. FPI was measured using Crystal Chem Mouse Insulin Elisa kits according to the manufacturer’s instructions. HOMA-IR was calculated as:
8
Glucose and Insulin Tolerance Assessments
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Glucose tolerance was assessed at 3–4 weeks after surgery by injecting α-d -glucose (1.5 mg/kg, 30% w/v in sterile saline, i.p.) and measuring blood glucose from the tail vein before and at 15, 30, 60, and 120 min after injection, with glucose strips and a glucometer (Onetouch Ultra Strips and Onetouch Ultra Glucometer, LifeScan INC, Milpitas, CA). Glucose tolerance tests were conducted between 09:00 and noon, after 3–5 h of food deprivation.
Insulin tolerance was assessed at 9–10 weeks after surgery by injecting insulin (0.6 U/kg in sterile saline, i.p., Novolin R, Novo Nordisk, Bagsvaerd, Denmark) and measuring blood glucose as above.
Insulin tolerance was assessed at 9–10 weeks after surgery by injecting insulin (0.6 U/kg in sterile saline, i.p., Novolin R, Novo Nordisk, Bagsvaerd, Denmark) and measuring blood glucose as above.
9
Glucose and Insulin Tolerance Testing
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Glucose tolerance was assessed at 30 ± 4 days (referred to as 4 weeks) after surgery by injecting α-D-glucose (1.5 mg/kg, 30% w/v in sterile saline, i.p.) and measuring blood glucose from the tail vein before and at 15, 30, 60, and 120 min after injection, with glucose strips and a glucometer (Onetouch Ultra Strips and Onetouch Ultra Glucometer, LifeScan INC, Milpitas, CA, USA). Glucose tolerance tests were conducted between 09:00 and noon, after 3–5 h of food deprivation. Insulin tolerance was assessed at 43 ± 4 days (referred to as 6 weeks) after surgery by injecting insulin (0.6 U/kg in sterile saline, i.p., Novolin R, Novo Nordisk, Bagsvaerd, Denmark) and measuring blood glucose as above.
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