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Ultrasensitive mouse insulin kit

Manufactured by Crystal Chem
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The Ultrasensitive mouse insulin kit is a laboratory equipment designed to measure insulin levels in mouse samples. It provides a sensitive and accurate method for quantifying insulin concentrations.

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Lab products found in correlation

Glucose oxidase method, by Applygen (2 mentions) Accu check performa, by Roche (2 mentions) Tissue triglyceride assay kit, by Applygen (1 mentions) Novolin 30r, by Novo Nordisk (1 mentions) Human insulin, by Merck Group (1 mentions) D12492, by Research Diets (1 mentions) Comprehensive lab animal monitoring system clams, by Columbus Instruments (1 mentions) Contour blood glucose monitor and strips, by Bayer (1 mentions) Mje00, by R&D Systems (1 mentions) Elisa kit, by Cell Sciences (1 mentions) Elisa kit, by R&D Systems (1 mentions) Freestyle glucometer, by Abbott (1 mentions) Fluorimetric assay kit, by Cayman Chemical (1 mentions) Mouse elisa kit, by R&D Systems (1 mentions)

10 protocols using ultrasensitive mouse insulin kit

1

Serum Biomarkers Analysis Protocol

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Serum glucose level was analyzed using glucose oxidase method (Applygen, Beijing, China) (Wang C. et al., 2016 (link)). Serum insulin level was determined by ELISA using an ultra-sensitive mouse insulin kit (Crystal Chem, USA) (Ding et al., 2016 (link)). Serum levels of total cholesterol, triglycerides, LDL-C, HDL-C, and free fatty acids (FFA) levels were assayed using the calorimetric kits from Nanjing Jiancheng Bioengineering Institute (Nanjing, China) (Jiang et al., 2016 (link); Wang et al., 2019 (link); Liu et al., 2020 (link)). Liver TG was analyzed using Tissue triglyceride assay kit (Applygen, Beijing, China) (Wang C. et al., 2016 (link)).
Guo F., Xu S., Zhu Y., Zheng X., Lu Y., Tu J., He Y., Jin L, & Li Y. (2020). PPARγ Transcription Deficiency Exacerbates High-Fat Diet-Induced Adipocyte Hypertrophy and Insulin Resistance in Mice. Frontiers in Pharmacology, 11, 1285.
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2

Glucose and Insulin Tolerance in Mice

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Mice treated with drugs were fasted for 6 h with free access to water. For the glucose tolerance test (GTT), 1 g/kg of glucose was i.p. injected into the mice, and blood glucose was measured with the Accu-Check® Performa (Roche Applied Science, Germany) at 0, 30, 60, 90, and 120 min. For the insulin tolerance test (ITT), 1 units/kg of recombinant human insulin (Novolin 30R, Novo Nordisk, Denmark) was i.p. injected into the mice, and blood glucose was measured at 0, 30, 60, 90, and 120 min after insulin injection. Serum glucose was determined using the Glucose Oxidase Method (APPLYGEN, China), and serum insulin levels were determined by ELISA using an ultra-sensitive mouse insulin kit (Crystal Chem, USA). The cholesterol, Hb1Ac, GSP and ALT levels were assayed using the kits from Nanjing Jiancheng Bioengineering Institute (Nanjing, China).
Zheng W., Qiu L., Wang R., Feng X., Han Y., Zhu Y., Chen D., Liu Y., Jin L, & Li Y. (2015). Selective targeting of PPARγ by the natural product chelerythrine with a unique binding mode and improved antidiabetic potency. Scientific Reports, 5, 12222.
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3

Metabolic Profiling of TACI-Deficient Mice

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Wild-type and TACI KO female mice were fed either normal chow diet (NCD) (6% fat) or HFD (D12492, 60% kcal fat; Research Diets, New Brunswick, NJ). After 14 weeks, fasting insulin levels were measured with an ultrasensitive mouse insulin kit (Crystal Chem Inc, Downers Grove, IL). For glucose tolerance tests (GTTs), fasted mice were i.p. injected with 1.5 g/kg glucose. For insulin tolerance test (ITTs), mice received 0.5 units/kg human insulin (Sigma-Aldrich, St. Louis, MO), after which blood glucose was measured using the OneTouch Ultra 2 glucose monitoring system (LifeScan, Milpitas, CA). Lean and fat mass compositions were determined by DEXA using a GE Lunar PIXImus densitometer (GE Lunar Corp.). For measurement of VO2, food intake, and locomotor activity, mice were analyzed using Comprehensive Lab Animal Monitoring System (CLAMS) (Columbus Instruments, Columbus, OH). Mice were allowed to acclimatize for 12 h before measurements were taken. VO2, VCO2, and heat measurements were normalized to lean mass or lean and fat mass combined as determined by DEXA analysis at the end of the CLAMS experiment. Subcutaneous AT (SAT) of inguinal fat pad and visceral AT (VAT) of perigonadal fat pad, brown fat pad, and liver of mice were dissected and weighed after euthanasia.
Liu L., Inouye K.E., Allman W.R., Coleman A.S., Siddiqui S., Hotamisligil G.S, & Akkoyunlu M. (2018). TACI-Deficient Macrophages Protect Mice Against Metaflammation and Obesity-Induced Dysregulation of Glucose Homeostasis. Diabetes, 67(8), 1589-1603.
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4

Glucose Tolerance Test in Mice

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Intraperitoneal (i.p.) GTTs were performed on weeks 3, 6, and 9 on non-anesthetized mice. Mice were fasted for 8 h and given an i.p. injection of glucose (i.e., 20% solution at 1 g/kg body weight). Blood from the tail vein collected at baseline and 5, 15, 30, 60, and 120 minutes post-i.p. glucose injection was used to quantify glucose levels using a Bayer Contour blood glucose monitor and strips (Bayer Healthcare, Tarrytown, NY, USA). Plasma insulin levels were detected using an ultrasensitive mouse insulin kit (Crystal Chem, Inc, Downers Grove, IL). The homeostasis model assessment method (HOMA) for insulin resistance (IR) was used employing the following formula: [fasting insulin concentration (ng/ml) × 24 × fasting glucose concentration (mg/dl)]/405 [16 ].
Baldwin J., Collins B., Wolf P.G., Martinez K., Shen W., Chuang C.C., Zhong W., Cooney P., Cockrell C., Chang E., Gaskins H.R, & McIntosh M.K. (2015). Table grape consumption reduces adiposity and markers of hepatic lipogenesis and alters gut microbiota in butter fat-fed mice. The Journal of nutritional biochemistry, 27, 123-135.
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5

Glucose Tolerance and Inflammation Markers

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Intraperitoneal (i.p.) GTTs were performed on weeks 7, 12, and 16 on non-anesthetized, fasted mice as previously described [33 ]. Plasma insulin levels were detected using an ultrasensitive mouse insulin kit (Crystal Chem, Inc, Downers Grove, IL). The homeostasis model of assessment (HOMA) for insulin resistance (IR) was calculated as described [32 (link), 33 ].
Plasma monocyte chemoattractant protein (MCP)-1 was measured using an ELISA kit from R&D Systems, Minneapolis, MN, USA (#MJE00). Plasma lipopolysaccharide (LPS) binding protein (LBP), an acute-phase protein that initiates recognition of LPS and activates host immune responses, was determined using an ELISA kit from Cell Sciences, Canton, MA (#CKM043).
Collins B., Hoffman J., Martinez K., Grace M., Lila M.A., Cockrell C., Nadimpalli A., Chang E., Chuang C.C., Zhong W., Mackert J., Shen W., Cooney P., Hopkins R, & McIntosh M. (2016). A Polyphenol-Rich Fraction Obtained from Table Grapes Decreases Adiposity, Insulin Resistance, and Markers of Inflammation and Impacts Gut Microbiota in High-Fat Fed Mice. The Journal of nutritional biochemistry, 31, 150-165.
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6

Metabolic Profile Measurement Protocol

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Insulin levels were measured in serum using the Ultra Sensitive Mouse Insulin kit from Crystal Chem (Zaandam, Netherlands), leptin and adiponectin levels were measured in plasma using the mouse ELISA kits from R&D Systems (Abingdon, UK), total cholesterol was measured using the fluorimetric assay kit from Cayman Chemicals via Biomol (Hamburg, Germany) and triglycerides were measured using the Fluitest TG kit from Analyticon (Lichtenfels, Germany) all according the manufacturer’s instructions. Glucose levels in whole blood were measured using the Free Style glucometer from Abbott (Abbott Park, Illinois, United States).
Pallauf K., Chin D., Günther I., Birringer M., Lüersen K., Schultheiß G., Vieten S., Krauß J., Bracher F., Danylec N., Soukup S.T., Kulling S.E, & Rimbach G. (2019). Resveratrol, lunularin and dihydroresveratrol do not act as caloric restriction mimetics when administered intraperitoneally in mice. Scientific Reports, 9, 4445.
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7

Glucose Tolerance Test in Mice

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Before the oral glucose tolerance test (OGTT), mice were fasted for 6 h with free access to water. Then, the mice were oral gavaged with 1 g/kg of glucose, and the levels of blood glucose were measured using an Accu-Check Performa (Roche Applied Science, Mannheim, Germany) at 0, 15, 30, 60, 90 and 120 min. Serum insulin levels were measured using an ultra-sensitive mouse insulin kit (Crystal Chem. Inc., Elk Grove Village, IL, USA).
Guo F., Zhu Y., Han Y., Feng X., Pan Z., He Y., Li Y, & Jin L. (2022). DEPP Deficiency Contributes to Browning of White Adipose Tissue. International Journal of Molecular Sciences, 23(12), 6563.
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8

Pancreas Insulin and Glucagon Extraction

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The whole pancreas was dissected free from fat immediately after the mouse was killed. After the measurement of the wet weight, the pancreas was homogenized with ULTRA-TURRAX and acid-ethanol extracted (0.18 M HCl in 70% ethanol) for 16 h at 4 °C. Insulin and glucagon concentration in clarified supernatant (after centrifugation at 4000 g) were determined by Ultrasensitive Mouse Insulin kit (Crystal Chem).
Vauthier V., Roujeau C., Chen P., Sarkis C., Migrenne S., Hosoi T., Ozawa K., Rouillé Y., Foretz M., Mallet J., Launay J.M., Magnan C., Jockers R, & Dam J. (2016). Endospanin1 affects oppositely body weight regulation and glucose homeostasis by differentially regulating central leptin signaling. Molecular Metabolism, 6(1), 159-172.
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9

Glucose and L-Arginine Injection Protocols

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Male and female mice were fasted for 16 h and injected IP with 2 g/kg body weight of 20% glucose (SIGMA G7021) in PBS or 1 g/kg body weight of 10% L-Arginine (SIGMA A8094) in PBS. For insulin measurements, blood (50 ul) was collected in hepatized capillary tubes (FisherBrand 22-260-950) at the indicated times. Samples were clarified by centrifugation at 15,000×g for 3 min and stored at −80 °C prior to the assay with the Ultra-Sensitive Mouse Insulin Kit (Crystal Chem 90080) according to the manufacturer’s protocol.
Iorio C., Rourke J.L., Wells L., Sakamaki J.I., Moon E., Hu Q., Kin T, & Screaton R.A. (2021). Silencing the G-protein coupled receptor 3-salt inducible kinase 2 pathway promotes human β cell proliferation. Communications Biology, 4, 907.
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10

Fasting Insulin Peptide Levels

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Plasma insulin, proinsulin, and C-peptide levels in mice fasted for 16 h were determined using an ultrasensitive mouse insulin kit (Crystal Chem Inc., Downers Grove, IL) and mouse proinsulin and C-peptide ELISA kits (ALPCO Diagnostics, Salem, NH), respectively.
Li L.C., Wang Y., Carr R., Haddad C.S., Li Z., Qian L., Oberholzer J., Maker A.V., Wang Q, & Prabhakar B.S. (2014). IG20/MADD Plays a Critical Role in Glucose-Induced Insulin Secretion. Diabetes, 63(5), 1612-1623.
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