A glucose tolerance test was performed in overnight fasted mice by injecting D-glucose (2 g/kg body weight) intraperitoneally. Blood glucose concentrations were monitored by tail bleeding at 0, 15, 30, 60, and 120 min after the glycemic load. Blood glucose levels were measured using a glucometer (ACCU-CHEK, Roche, Indianapolis, IN, USA). Insulin levels were measured using an ultrasensitive insulin ELISA kit (#90060, Crystal Chem, Elk Grove Village, IL, USA) [65 (link)]. A subset of mice was used to examine tissue insulin sensitivity—mice were fasted for 16 h before i.p. injection of 2.5 U/kg insulin (NovoRapid, Novo Nordisk, Bagsværd, Denmark). After 5 min, mice were euthanized and tissues were harvested, shock frozen in liquid nitrogen, and stored for later assays.
Novorapid
Manufactured by Novo Nordisk
Sourced in Denmark, France, China
NovoRapid is a fast-acting insulin product used to control blood sugar levels in individuals with diabetes. It is a man-made version of the naturally occurring insulin hormone and is designed to be rapidly absorbed into the bloodstream to provide quick-acting blood sugar control. NovoRapid is administered via subcutaneous injection.
Automatically generated - may contain errors
Lab products found in correlation
D glucose, by Merck Group (3 mentions)
Humalog, by Eli Lilly (3 mentions)
Levemir, by Novo Nordisk (3 mentions)
Lantus, by Sanofi (3 mentions)
Immulite 1000 immunoassay system, by Siemens (2 mentions)
Accu check, by Roche (2 mentions)
Apidra, by Sanofi (2 mentions)
Tresiba, by Novo Nordisk (2 mentions)
Onetouch ultra glucometer, by LifeScan (2 mentions)
Novomix 30, by Novo Nordisk (2 mentions)
Glucagon, by Merck Group (2 mentions)
Onetouch vita, by LifeScan (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Glutamax, by Thermo Fisher Scientific (2 mentions)
Accu chek, by Roche (1 mentions)
Insulin elisa kit, by Crystal Chem (1 mentions)
Ripa lysis buffer, by Roche (1 mentions)
Protease and phosphatase inhibitor, by Roche (1 mentions)
D glucose, by Thermo Fisher Scientific (1 mentions)
77 protocols using novorapid
1
Glucose Tolerance Test in Fasted Mice
2
Palmitic Acid-Induced Insulin Resistance
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The treatment with palmitic acid (PA) was conducted on serum-starved cells as previously described [18 (link)]. Before the addition of palmitate to the medium, it was conjugated with fatty acid-free bovine serum albumin (BSA, Sigma-Aldrich, St. Louis, MO). Briefly, palmitate stock solution was prepared by dissolving PA in a mixture of absolute ethanol and 1 M NaOH, heating to 70°C and conjugating with 10% BSA. Next, stock solution was diluted in serum free-DMEM, containing 10mM Hepes. Next the cells were incubated in the presence or absence of the palmitic acid at the concentration of 0.75 mM for 16h and 40h. At the end of each experimental set cells morphology as well as viability was assessed (i.e. Trypan blue staining). In selected sets (as indicated in Figures) the hepatocytes were chased by 100nM insulin (NovoRapid, Novo Nordisk, Ontario, Canada) for 15 min at 37°C then washed three times with PBS, harvested and homogenized in ice cold RIPA lysis buffer containing protease and phosphatase inhibitors (Roche Diagnostics GmbH, Mannheim, Germany). Other sets of palmitate induced insulin resistant hepatocytes were transferred for further analysis of glycogen synthesis or sphingolipid content.
3
Insulin Sensitivity Assessment via Glucose Clamp
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Baseline samples were collected at −5 and −1 min before a bolus of glucose (50% dextrose; 11.4 g/m2 × body surface area) was infused intravenously over 60 s beginning at time 0. At 20 min, human insulin (0.02 U/kg; NovoRapid, Novo Nordisk) was infused over 5 min at a constant rate (HK400 Hawkmed Syringe Pump, Shenzhen Hawk Medical Instrument Co., Shenzhen, China) and samples were collected up to 240 min. Bergman’s minimal model of glucose kinetics was used to calculate the insulin sensitivity index (SI)36 (link). Samples for insulin (IMMULITE 1000 immunoassay system, Siemens Healthcare, Midrand, South Africa) and glucose (Randox, Gauteng, South Africa) were collected in serum-separating and fluoride oxalate tubes, respectively. Samples were centrifuged at 3000 rpm for 10 min at 4 °C and stored at −80 °C until further analyses.
4
Glucose Tolerance and Insulin Sensitivity Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After 14 weeks on the diets, mice underwent an intraperitoneal ITT. After 6 h of fasting, from 08:00 to 14:00, mice were injected with 0.75 U/kg body weight of human insulin (Novorapid, Novo Nordisk, Bagsværd, Denmark). Blood glucose levels were measured at 0, 20, 40, 60, 90 and 120 min. After 15 weeks on the diets, mice underwent an IPGTT. After an overnight fast, from 18:00 to 09:00, mice were injected with 2 g/kg body weight of d -glucose (Invitrogen, Bleiswijk, the Netherlands). Blood glucose was measured at 0, 15, 30, 60 and 120 min. See ESM for full methods.
5
Glucose Metabolism Evaluation in Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
GTT and ITT experiments were conducted to evaluate the glucose metabolic rate of the correspondingly treated mice. For the GTT, mice were fasted overnight (from 5 p.m. to 9 a.m.), and fasting blood glucose was assessed (0 min). Then, 2 g/kg glucose was injected intraperitoneally (i.p.), and tail blood glucose was measured using a handheld glucometer (Ascensia Breeze, Bayer Company, Germany) at 15, 30, 60, 90, 120, and 150 min after glucose injection. For the ITT, mice were fasted for 4 h with free access to drinking water and the basal blood glucose levels were recorded (0 min), after which the mice received an i.p. injection of 0.75 units/kg insulin (NovoRapid, Novo Nordisk); and the glucose concentrations were determined at 15, 30, 60, and 90 min after insulin injection.
6
Insulin Glucose Tolerance Test
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Male C57BL/6J or Tst—/— mice were maintained on standard chow (RM1). Mice were fasted for 4 hours prior to injection i.p. of insulin (1 mU/g bodyweight, NovoRapid 100U/ml, Novo Nordisk). Tail venesection blood samples were taken prior to, and 15, 30, 60 and 120 minutes post injection. Blood glucose was measured from samples using a Glucometer (Accu-Check, Performa Nano, Roche). Blood glucose was plotted across time to evaluate net glucose accumulation in blood.
7
Normoglycemic Pig Glucagon Study
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The pigs received separate continuous IV infusions of 0.05 IU/kg/hour insulin aspart (NovoRapid®, Novo Nordisk AS, Denmark) and 20% glucose solution (Glucos B. Braun®, Braun, Germany) throughout the study day to prevent glycogen depletion. Before each glucagon bolus, blood glucose concentration was titrated to a normoglycaemic target concentration of 4–5 mmol/L by adjusting the glucose infusion rate. According to the study protocol, the blood glucose concentration had to be stable (no more than 0.2 mmol/L variations) before every glucagon administration, without any glucose infusion rate adjustments for 20 min. The glucose infusion rate was also kept stable for the first 60 min after glucagon administration to monitor the effects on glucose metabolism.
8
Two-Step Hyperinsulinemic-Euglycemic Clamp
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After an overnight fast of at least 10 hours, participants were admitted to the Clinical Research Unit of the VU University Medical Center at 08:30 hours. First, body weight, body fat percentage, hip/waist circumference, and blood pressure were measured. Next, insulin sensitivity was assessed by a 2-step hyperinsulinemic-euglycemic clamp as previously described. 16 (link) Briefly, a primed (500 mU•m -2 ) continuous insulin (NovoRapid, Novo Nordisk, Bagsvaerd, Denmark) infusion was started at a rate of 20 mU•m -2 •min -1 (step 1) for the total duration of 120 minutes and raised to 60 mU•m -2 •min -1 (step 2) for another 120 minutes. Venous glucose concentrations were measured every 5 minutes at bedside (YSI 2300 STAT Plus Analyzer, Yellow Springs, OH), and glucose 20% was infused to maintain a plasma glucose concentration of 5 mmol/L. Peripheral GU (M value) was calculated as the glucose infusion rate at steadystate (last 60 minutes) during both steps. At baseline, at the end of step 1 and at the end of step 2, 3 blood samples were drawn among others for the determination of insulin and FFA concentrations. Before and during the last hour of the 2-step clamp, we performed indirect calorimetry, contrast-enhanced ultrasonography, and continuous peripheral plethysmography (Figure 1). Additional information on the performed test can be found in the Data Supplement.
9
Measuring Glucose Homeostasis in Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Blood glucose levels were verified by intraperitoneal glucose tolerance test (IPGTT) and insulin tolerance test (ITT), as previously described [41] (link). Briefly, the IPGTT was performed in mice fasted for 6 hours, administering intraperitoneally (i.p.) glucose at 1 g/Kg body weight.
Subsequently, blood samples were collected from the tail vein at 0, 30, 60, 90 and 120 min after glucose injection. Fasting blood glucose (FBG) levels were measured with an automatic glucometer (Glucomen LX, Menarini Diagnostics, Firenze). Likewise, the ITT was carried out after i.p. injection of insulin (1Units/kg body weight Regularâ, Novorapid, Novonordisk, Rome, Italy), as previously described [43] (link). Briefly, blood samples were collected from the tail vein at 0, 30, 60, 90 and 120 min and FBG levels were assessed as reported above.
Subsequently, blood samples were collected from the tail vein at 0, 30, 60, 90 and 120 min after glucose injection. Fasting blood glucose (FBG) levels were measured with an automatic glucometer (Glucomen LX, Menarini Diagnostics, Firenze). Likewise, the ITT was carried out after i.p. injection of insulin (1Units/kg body weight Regularâ, Novorapid, Novonordisk, Rome, Italy), as previously described [43] (link). Briefly, blood samples were collected from the tail vein at 0, 30, 60, 90 and 120 min and FBG levels were assessed as reported above.
10
Metabolic Assessment in Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Body composition was measured by MRI using an EchoMRI (Echo Medical Systems). Wholebody insulin sensitivity was assessed after the indicated time on CD or HFD in 4 h fasted mice by an i.p. insulin tolerance test (ipITT). After an initial blood collection (t=0), an i.p. bolus of insulin (1 U/kg lean body mass; NOVORAPID, Novo Nordisk) was administered to the mice.
Blood glucose was measured by tail bleeding at 15, 30, 60, and 120 min after insulin administration using a Glucometer. Whole-body glucose tolerance was assessed after the indicated time on CD or HFD in 6 h fasted mice by an intraperitoneal glucose tolerance test (ipGTT). After an initial blood collection (t=0), an i.p. injection of glucose (2g D-Glucose/kg total body weight, Sigma-Aldrich) was administered in conscious mice. Blood glucose was measured by tail bleeding at 15, 30, 60 and 120 min after glucose administration using a Glucometer (Accu-Check, Roche Diagnostics). At 15 minutes, blood was also collected for analysis of plasma insulin levels as described above.
Blood glucose was measured by tail bleeding at 15, 30, 60, and 120 min after insulin administration using a Glucometer. Whole-body glucose tolerance was assessed after the indicated time on CD or HFD in 6 h fasted mice by an intraperitoneal glucose tolerance test (ipGTT). After an initial blood collection (t=0), an i.p. injection of glucose (2g D-Glucose/kg total body weight, Sigma-Aldrich) was administered in conscious mice. Blood glucose was measured by tail bleeding at 15, 30, 60 and 120 min after glucose administration using a Glucometer (Accu-Check, Roche Diagnostics). At 15 minutes, blood was also collected for analysis of plasma insulin levels as described above.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!