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Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary: Triglyceride emulsion and heparin will be intravenously infused for 5 hours to acutely raise circulating fatty acids levels in awake mice. Acute lipid infusion is shown to cause insulin resistance in peripheral organs.

Proper citation: Jason Kim 2019. U Mass - Acute lipid infusion. protocols.io https://dx.doi.org/10.17504/protocols.io.xm5fk86 Copy   

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Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary: This experiment involves a spectrophotometric measurement using Roche Cobas Clinical Chemistry Analyzer.

Proper citation: Jason Kim 2019. U Mass - Lactate dehydrogenase. protocols.io https://dx.doi.org/10.17504/protocols.io.x2yfqfw Copy   

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Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary: Summary: Insulin clearance test measure systemic clearance of insulin following a bolus injection. Liver accounts for the majority of systemic insulin clearance following secretion from pancreatic β- cells into portal circulation. Hepatic clearance of insulin may be affected by obesity and in other mouse models of altered metabolism. Alterations in insulin clearance may also affect glucose and lipid metabolism.

Proper citation: Jason Kim 2019. U Mass - Insulin clearance. protocols.io https://dx.doi.org/10.17504/protocols.io.x2wfqfe Copy   

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Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary: This experiment involves a spectrophotometric measurement using Roche Cobas Clinical Chemistry Analyzer. Serum lactate levels are affected by alterations in glucose and protein metabolism.

Proper citation: Jason Kim 2019. U Mass -Lactate. protocols.io https://dx.doi.org/10.17504/protocols.io.x5ufq6w Copy   

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Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary: Summary:This experiment provides the quantification of multiple hormones using multiplexed-Luminex technology based on beads containing specific antibodies. Sample matrices include plasma, serum, tissue culture supernatants, and tissue or cell lysates. Serum C-peptide levels reflect insulin secretion and pancreatic -cell function. Serum C-peptide levels are altered in obesity, insulin resistance, and type 1 and type 2 diabetes.

Proper citation: Jason Kim 2019. U Mass - C-Peptide. protocols.io https://dx.doi.org/10.17504/protocols.io.xttfnnn Copy   

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Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary: Summary: This experiment provides the quantification of multiple cytokines and chemokines using multiplexed-Luminex technology based on beads containing specific antibodies. Serum cytokine levels reflect chronic or acute inflammation, and circulating cytokines and chemokines are altered in obesity. Cytokines Panel III include IL-20 (interleukin-20), IL-23, IL-27, IL-33, MDC (macrophage- derived chemokine; CCL-22), and TIMP-1 (tissue inhibitor of metalloproteinases). A service can be requested for all or any combination of listed cytokines/chemokines for customized multiplexed Luminex assay.

Proper citation: Jason Kim 2019. U Mass - Cytokines Panel III - multiplex. protocols.io https://dx.doi.org/10.17504/protocols.io.xv7fn9n Copy   

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Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary: Summary: This experiment provides the quantification of multiple cytokines and chemokines using multiplexed-Luminex technology based on beads containing specific antibodies. Serum cytokine levels reflect chronic or acute inflammation, and circulating cytokines and chemokines are altered in obesity. Cytokines Panel II include IL-16 (interleukin-16), IL-17E/IL-25, IL-21, IL-22, IL-28B, EPO (erythropoietin), Exodus-2 (CCL-21), Fractalkine (CX3CL1), MCP-5 (monocyte chemotactic protein-5; CCL-12), MIP-3α (macrophage inflammatory protein 3-alpha; CCL-20), MIP-3β(macrophage inflammatory protein 3-beta; CCL-19), and TARC (thymus and activation- regulated; CCL-17). A service can be requested for all or any combination of listed cytokines/chemokines for customized multiplexed Luminex assay.

Proper citation: Jason Kim 2019. U Mass - Cytokines Panel II - multiplex. protocols.io https://dx.doi.org/10.17504/protocols.io.xvvfn66 Copy   

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Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary: A high-fat diet of varying composition and percent fat is administered to induce obesity in mice. High-fat diet induced obesity is causally associated with insulin resistance and type 2 diabetes.

Proper citation: Jason Kim 2019. U Mass - Chronic high-fat feeding. protocols.io https://dx.doi.org/10.17504/protocols.io.xugfntw Copy   

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Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary: Summary:The TSE PhenoMaster/LabMaster Metabolic Cage system is used to measure indirect calorimetry, food/water intake, energy expenditure, and physical activity in individually housed awake mice. The experiment noninvasively measures VO² consumption and VCO² production rates in individual mice using metabolic chambers and calculates the respiratory exchange ratio (respiratory quotient) to reflect energy expenditure. Metabolic cage measurement is conducted continuously for 72 hours (3 days) to account for acclimation of mice housed in home cages during the study. With our high-speed Siemens O²/CO² sensing unit, indirect calorimetry measurements may be performed at a 20-min interval for a full 12-cage study.

Proper citation: Jason Kim 2019. U Mass - Energy balance – food intake, energy expenditure, physical activity. protocols.io https://dx.doi.org/10.17504/protocols.io.xwwfpfe Copy   

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Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary: Summary: This experiment provides the quantification of multiple cytokines and chemokines using multiplexed-Luminex technology based on beads containing specific antibodies. Serum cytokine levels reflect chronic or acute inflammation, and circulating cytokines and chemokines are altered in obesity. Cytokines Panel I include IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, KC (IL-8 homologue), IL-9, IL-10 (interleukin-10), IL-12 (p40), IL-12 (p70), IL-13, IL-15, IL-17A, TNFα (tumor necrosis factor alpha), IFNγ (interferon gamma), IP-10 (interferon gamma-induced protein 10; CXCL-10), Eotaxin (CCL-11), G-CSF (granulocyte colony stimulating factor), GM-CSF (granulocyte macrophage colony stimulating factor), LIF (leukemia inhibitory factor), LIX (LPS- induced CXC chemokine), MCP-1 (monocyte chemotactic protein-1; CCL-2), M-CSF (macrophage colony stimulating factor), MIG (monokine induced by gamma interferon; CXCL- 9), MIP-1α (macrophage inflammatory protein alpha; CCL-3), MIP-1β (macrophage inflammatory protein beta; CCL-4), and MIP-2/RANTES (macrophage inflammatory protein 2- alpha; CXCL-2). A service can be requested for all or any combination of listed cytokines/chemokines for customized multiplexed Luminex assay.

Proper citation: Jason Kim 2019. U Mass - Electrolytes. protocols.io https://dx.doi.org/10.17504/protocols.io.xwafpae Copy   

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Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary: This experiment involves a spectrophotometric measurement using Roche Cobas Clinical Chemistry Analyzer.

Proper citation: Jason Kim 2019. U Mass - Creatine Kinase. protocols.io https://dx.doi.org/10.17504/protocols.io.xumfnu6 Copy   

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Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary: Summary:The insulin enzyme-linked immunosorbent assay (ELISA) is a standard immunoassay for the detection of insulin levels in serum samples. This experiment uses a 96-well plate coated with anti-insulin antibodies and developed with a secondary anti-insulin antibody linked to horseradish peroxidase. The ELISA is developed with 3,3',5,5'-Tetramethylbenzidine (TMB), a chromogenic substrate and read on an automated plate spectrophotometer at 450 nm. Insulin ELISA may be processed with normal or ultra-sensitive assay kit depending on the expected range of sample concentrations. Serum insulin levels are affected by pancreatic β-cell function and insulin sensitivity. Serum insulin levels are elevated in obesity and insulin resistance.

Proper citation: Jason Kim 2019. U Mass - Insulin. protocols.io https://dx.doi.org/10.17504/protocols.io.xznfp5e Copy   

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Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary: Insulin tolerance test measures systemic clearance of glucose following an intraperitoneal bolus injection of a physiological dose of insulin. This experiment measures insulin sensitivity in awake mice assuming that there are no alterations in the animal’s counterregulatory response. Insulin sensitivity is altered in obese mice.

Proper citation: Jason Kim 2019. U Mass - Insulin tolerance test. protocols.io https://dx.doi.org/10.17504/protocols.io.x2xfqfn Copy   

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Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary: Summary:Hyperinsulinemic-euglycemic clamp is the gold-standard method to assess insulin sensitivity. The hyperinsulinemic-euglycemic clamp is widely used in clinics and laboratories to measure insulin action on glucose utilization in humans and animals for clinical and basic science research. Incorporation of radioactive-labeled glucose during hyperinsulinemic-euglycemic clamps makes it possible to measure glucose metabolism in individual organs in awake mice. Impaired insulin sensitivity (insulin resistance) is a major characteristic of obesity and an early requisite event in the development of type 2 diabetes.

Proper citation: Jason Kim 2019. U Mass - Hyperinsulinemic-euglycemic clamp. protocols.io https://dx.doi.org/10.17504/protocols.io.xzefp3e Copy   

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Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary: Intravenous administration via tail vein is used to acutely deliver drug, hormones, and adenoassociated virus in mice. A large fraction of injectate will be cleared by liver.

Proper citation: Jason Kim 2019. U Mass - Surgery – tail vein injection. protocols.io https://dx.doi.org/10.17504/protocols.io.x5dfq26 Copy   

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Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary: Chronic indwelling catheter is placed in the carotid artery for blood sampling during experiments. Surival surgery is performed in anesthetized mice, and mice recover from surgery after 4~5 days.

Proper citation: Jason Kim 2019. U Mass - Surgery – carotid artery cannulation. protocols.io https://dx.doi.org/10.17504/protocols.io.x4pfqvn Copy   

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Authors: Jason Kim
Group: Mouse Metabolic Phenotyping Centers
Summary: This experiment measures blood or plasma glucose concentration using Analox GM9 Glucose Analyzer or GM7 Micro-Stat Rapid Multi-assay Analyzer. The measurement is based on the rate of oxygen uptake in the reaction between sample glucose and glucose oxidase. Plasma glucose levels are elevated (i.e., hyperglycemia) in obesity, insulin resistance, and type 2 diabetes.

Proper citation: Jason Kim 2019. U Mass - Glucose. protocols.io https://dx.doi.org/10.17504/protocols.io.xw5fpg6 Copy   

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