Humulin r u 100

Manufactured by Eli Lilly

Sourced in United States

Humulin R U-100 is an injectable human insulin solution used for the treatment of diabetes mellitus. It is a short-acting insulin that helps regulate blood sugar levels. The product is available in a 100 units per milliliter concentration.

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24 protocols using humulin r u 100

Insulin tolerance test in mice

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Mice were fasted from 9 am to 3 pm. Following fasting, mice were injected intraperitoneally with 0.375 U/Kg of insulin (Humulin R U-100 Lilly) in PBS and tail blood glucose levels were measured at 15, 30, 45, 60, 90, and 120 min after injection. Glucose was monitored with Accu-check Aviva glucometer. In case of the appearance of seizures due to extremely low levels of glucose, mice were injected with 30% glucose (Sigma #G7528) in PBS.

Plata-Gómez A.B., de Prado-Rivas L., Sanz A., Deleyto-Seldas N., García F., de la Calle Arregui C., Silva C., Caleiras E., Graña-Castro O., Piñeiro-Yáñez E., Krebs J., Leiva-Vega L., Muñoz J., Jain A., Sabio G, & Efeyan A. (2024). Hepatic nutrient and hormone signaling to mTORC1 instructs the postnatal metabolic zonation of the liver. Nature Communications, 15, 1878.

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Dietary FA Composition Analysis

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Nutrients and other chemical compounds, vitamins and minerals for the diet preparations, were of chemical grade or better, with the exception of corn oil (Arcor, Córdoba, Argentina), hydrogenated coconut oil without TFA (Castoroil, Buenos Aires, Argentina), partially hydrogenated soybean oil (-50% of TFA-Calsa, Buenos Aires, Argentina), sucrose, cellulose, and corn starch, which were obtained from local sources. Corn oil was used as an unsaturated cis-FA source, rich in LA (51% of total FA to resemble the FA composition of the Western diet). Coconut oil was used to produce a LA deficiency status and partially hydrogenated soybean oil was used as a TFA source. All solvents and reagents used for the FA quantification were of chromatography grade, and all other chemicals used were of at least American Chemical Society grade. The standard FA were purchased from Sigma Chemical Co. (St Louis, MO, USA). The glucose test kits were commercially obtained from the Sociedad de Bioquímicos (Santa Fe, Argentina). The insulin used was Humulin R (U-100), which was acquired from Eli Lilly (Indianapolis, IN, USA). Amersham International (Bucks, UK) provided the 2-deoxy-[2,6-3 H]d-glucose and [U 14 C]d-glucose.

Fariña A.C., Hirabara S., Sain J., González M., Curi R, & Bernal C. (2018). Influence of trans fatty acids on glucose metabolism in soleus muscle of rats fed diets enriched in or deprived of linoleic acid. European journal of nutrition, 57(4).

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Metabolic Phenotyping in Mice

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Glucose and insulin tolerance test: Glucose and insulin tolerance tests were performed in 12 mos mice, following 10 mos on the WD. After fasting for 5 hours, glucose tolerance test (GTT) was performed (Agri Laboratories, LTD). For the insulin tolerance test (ITT), mice received a single intraperitoneal injection of a diluted insulin solution (males 0.75 IU/kg and females 0.50 IU/kg; prepared in sterile saline) (Humulin R U-100, Eli Lilly). Glucose was determined using a handheld glucometer (Bayer Contour Next).
NMR Body Composition: Body composition was determined in the Echo MRI 3-in-1, time domain nuclear magnetic resonance (TD-NMR) system (Houston, TX). Body weight was measured on a standard laboratory balance before mice were placed in to a clear, cylindrical holder. The tube was gently inserted in to the boor for an approximately two-minute measurement. Adiposity (% body fat) was calculated as ((fat mass/total body weight) x 100).
Plasma lipid measurements: Blood was collected in K2 EDTA (1.0mg) microtainer tubes (BD) at harvest (non-fasted) and kept at room temperature for at least 30 minutes to prevent clotting and then centrifuged at 22°C for 15 minutes at 5000rpm. Plasma was carefully collected and aliquoted. Plasma was characterized on the Beckman Coulter AU680 chemistry analyzer.

Graham L., Kocalis H., Soto I., & Howell G.R. (2020). Deficiency of complement component C1Q prevents cerebrovascular damage and white matter loss in a mouse model of chronic obesity.

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Insulin Tolerance and Glucose Tolerance in Mice

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ITT was performed in 18-week-old animals from setting #1 fasted for 6 hours. Blood glucose was measured at 0, 15, 30, 45, and 60 minutes from tail tip with an Accu-Check Active glucometer (Roche) after an i.p. injection of 0.75 U/kg insulin (Humulin R U-100, Lilly). GTT was performed in obese mice from setting #2, receiving LY or vehicle during 5 weeks, and fasted for 12 hours before the assay. There, blood glucose was measured at 0, 15, 30, 60, and 120 minutes after an i.p. injection of a 20% glucose solution (Glucocemin 33%, B. Braun). The areas under the glucose curves were evaluated according to the trapezoidal rule in each animal.

Lluch A., Veiga S.R., Latorre J., Moreno-Navarrete J.M., Bonifaci N., Dien Nguyen V., Zhou Y., Höring M., Liebisch G., Olkkonen V.M., Llobet-Navas D., Thomas G., Rodríguez-Barrueco R., Fernández-Real J.M., Kozma S.C, & Ortega F.J. (2022). A compound directed against S6K1 hampers fat mass expansion and mitigates diet-induced hepatosteatosis. JCI Insight, 7(14), e150461.

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Glucose and Insulin Tolerance Tests

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Mice were fasted for 6 h (GTT) or 4 h (ITT) followed by being administrated with 1.5 g/kg glucose (Acros organics, Fair Lawn, NJ) or 0.8 mU/kg insulin (Humulin® R U100, Eli Lilly and Company, Indianapolis, IN) via intraperitoneal injection, respectively. Tail vein blood at baseline and indicated time points after the injection was collected and measured for the glucose level by using handheld glucometer (Germaine™ Laboratories, San Antonio, TX).
For the GSIS assay, mice were fasted for 4 h, and then intraperitoneally injected with 1.5 g/kg glucose. Blood was collected via cheek punch at baseline and 15 minutes post glucose administration. Plasma samples were then isolated and tested for the insulin concentration by applying an ultra-sensitive mouse insulin ELISA kit (Crystal Chem, Elk Grove Village, IL).

Wang L., Wang B., Gasek N.S., Zhou Y., Cohn R.L., Martin D.E., Zuo W., Flynn W.F., Guo C., Jellison E.R., Kim T., Prata L.G., Palmer A.K., Li M., Inman C.L., Barber L.S., Al-Naggar I.M., Zhou Y., Kuchel G.A., Meves A., Tchkonia T., Kirkland J.L., Robson P, & Xu M. (2021). Targeting p21Cip1-Highly-Expressing Cells in Adipose Tissue Alleviates Insulin Resistance in Obesity. Cell metabolism, 34(1), 75-89.e8.

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Insulin Tolerance Test in Mice

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Animals were fasted for 6 h prior to administration of the insulin bolus. Cages were changed at the time of fasting. Vehicle and test compounds were administered 1 h prior to the insulin bolus. At t = 0, mice received a bolus of human insulin (ob/ob mice received an insulin bolus of 1.5U/kg; lean control mice received an insulin bolus of 0.5U/kg. DIO mice received an insulin bolus of 1U/kg; lean chow fed control mice received an insulin bolus of 0.75U/kg (Humulin® R-U100; Eli Lilly)) by intra-peritoneal (i.p.) injection (5 ml/kg). Blood samples were collected from the tail vein and blood glucose was measured at time points −60, 0, 15, 30, 60 and 120 min after the insulin bolus. Mice were re-fed after the last blood sampling.

Raichur S., Brunner B., Bielohuby M., Hansen G., Pfenninger A., Wang B., Bruning J.C., Larsen P.J, & Tennagels N. (2019). The role of C16:0 ceramide in the development of obesity and type 2 diabetes: CerS6 inhibition as a novel therapeutic approach. Molecular Metabolism, 21, 36-50.

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Glucose and Insulin Tolerance Tests in Mice

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Intraperitoneal glucose tolerance tests (i.p. GTT) were performed as previously published (26 (link)). Mice were fasted for four hours and injected i.p. with 1.5 g/kg dextrose (D20W). A tail laceration was made, and blood glucose was measured on a glucometer in duplicate prior to dextrose administration (“0” minutes) and again 15, 30, 45, 60 and 120 min after dextrose administration. The insulin tolerance test (ITT) was performed similarly. Mice were fasted for four hours and injected i.p. with 1 unit/kg Humulin R U-100 (Eli Lilly, Indianapolis, IN). Blood glucose was measured from a tail vein laceration in duplicate on a glucometer prior to and at 15, 30, 45 and 60 minutes after insulin administration. Both GTT and ITT were performed in freely moving, conscious mice.

Beck J., da Silva Teixeira S., Harrison K., Phillips G., He Y, & Sisley S. (2022). Paraventricular Vitamin D Receptors Are Required for Glucose Tolerance in Males but Not Females. Frontiers in Endocrinology, 13, 869678.

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Metabolic Phenotyping of Mice

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Lipid tolerance tests were performed by fasting the mice for 6 hours starting at 8:00 am and then orally gavaging the mice with 5 μL/g extra virgin olive oil. Plasma samples were collected over the next 4 hours for triglyceride measurements. Glucose and leucine tolerance tests were performed by fasting the mice for 6 hours starting at 8:30 am and then orally gavaging the mice with 1.5 mg/g D-glucose or 0.3 mg/g L-leucine respectively. Blood glucose was measured at regular intervals over the next 2 hours. Plasma samples were obtained at 0, 7, 15, and 60 minutes post-gavage for insulin measurements. Insulin tolerance tests were performed by first fasting the mice for 6 hours starting at 8:30 am and then injecting insulin (Humulin R U-100, Lilly, #0002-8215-17) intraperitoneally. Blood glucose was measured at regular intervals over the next 2 hours.

Huynh F.K., Hu X., Lin Z., Johnson J.D, & Hirschey M.D. (2017). Loss of sirtuin 4 leads to elevated glucose- and leucine-stimulated insulin levels and accelerated age-induced insulin resistance in multiple murine genetic backgrounds. Journal of inherited metabolic disease, 41(1), 59-72.

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Glucose and Insulin Tolerance Tests

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Glucose and insulin tolerance tests were performed on 6 month-old mice that were fasted for 6 hours. Glucose concentrations were determined with an Accu-Chek advantage glucometer (Roche) in blood collected from the tail vein at indicated time point. Insulin (0.75 U/kg for HFD group mice and 0.375 U/kg for NCD group mice) or glucose (2g/kg) was intraperitoneal injected into mice. Insulin was Humulin R (U-100) from Lilly.

Tsai S.Y., Rodriguez A.A., Dastidar S.G., Del Greco E., Carr K.L., Sitzmann J.M., Academia E.C., Viray C.M., Martinez L.L., Kaplowitz B.S., Ashe T.D., La Spada A.R, & Kennedy B.K. (2016). Increased 4E-BP1 expression protects against diet-induced obesity and insulin resistance in male mice. Cell reports, 16(7), 1903-1914.

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Insulin Signaling Pathway Analysis

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Reagents were from the following manufacturers: ketamine (KetaSet^®), Xylazine (AnaSed^®), Medium 199, HBSS, EGTA, HEPES, PenStrep and Gentamycin (Life Technology), Collagen 4 (Worthington), Humulin^® R U-100 (Lilly), 8-(4-chlorophenylthio) (CPT)-cAMP, dexamethasone, cycloheximide, bovine serum albumin, d-glucose and sodium pyruvate (Sigma–Aldrich), Lipofectamine2000 (Thermo Fisher), miRCURY LNA (Qiagen) (biotinylated mmu-205-5p and cel-39-3p), anti-FOXO1 (C29H4, # 2880S), anti-Phospho-FoxO1 (Thr24)/FoxO3a (Thr32) (#2599S), anti-Akt, anti-Phospho-Akt (Ser473) (D9E, #4060), anti-SHIP2 (#3397S) and anti-PTEN (#9188S) (Cell Signaling), anti-actin (ab8227) (Abcam).

Langlet F., Tarbier M., Haeusler R.A., Camastra S., Ferrannini E., Friedländer M.R, & Accili D. (2018). microRNA-205-5p is a modulator of insulin sensitivity that inhibits FOXO function. Molecular Metabolism, 17, 49-60.

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