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Name Authors DOI Group Summary Associated Publications RRIDs used Affiliations External URL Version Publication Date Proper Citation Record Last Update
UC Davis - Intravenous Glucose Tolerance Test
 
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Kent Lloyd, Kristin Grimsrud 10.17504/protocols.io.8i7huhn Mouse Metabolic Phenotyping Centers The intravenous glucose tolerance test is used to assess insulin sensitivity, glucose disposal and islet function in vivo by measuring glucose and insulin responses following intravenous glucose administration. University of California, Davis, University of California, Davis https://mmpc.org/shared/document.aspx?id=291&docType=Protocol 3 2019 Kent Lloyd, Kristin Grimsrud 2019. UC Davis - Intravenous Glucose Tolerance Test. protocols.io dx.doi.org/10.17504/protocols.io.8i7huhn 2021-03-29 03:14:47
U Michigan - Massons Trichrome staining
 
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Jeff Hodgin 10.17504/protocols.io.56ug9ew Mouse Metabolic Phenotyping Centers Massons Trichrome staining is used for detection of collagen fibers for tubulointerstitial fibrosis on formalin-fixed paraffin-embedded kidney tissue section. The collagen fibers will be stained blue, the nuclei black, and the background is stained red. University of Michigan - Ann Arbor https://mmpc.org/shared/document.aspx?id=311&docType=Protocol 2 2019 Jeff Hodgin 2019. U Michigan - Massons Trichrome staining. protocols.io dx.doi.org/10.17504/protocols.io.56ug9ew 2021-03-29 03:14:56
U Mass - Aspartate Transferase
 
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Jason Kim 10.17504/protocols.io.xr5fm86 Mouse Metabolic Phenotyping Centers This experiment involves a spectrophotometric measurement using Roche Cobas Clinical Chemistry Analyzer. Serum levels of aspartate transferase reflect liver function. University of Massachusetts https://mmpc.org/shared/document.aspx?id=181&docType=Protocol 1 2019 Jason Kim 2019. U Mass - Aspartate Transferase. protocols.io dx.doi.org/10.17504/protocols.io.xr5fm86 2021-03-29 03:14:56
U Mass - Leptin
 
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Jason Kim 10.17504/protocols.io.x23fqgn Mouse Metabolic Phenotyping Centers 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. Leptin is an adipocyte-derived hormone (adipokine) that regulates feeding behavior (hunger/satiety). Serum leptin levels are altered in obesity, insulin resistance, and type 2 diabetes. University of Massachusetts http://mmpc.org/shared/document.aspx?id=164&docType=Protocol 1 2019 Jason Kim 2019. U Mass - Leptin. protocols.io dx.doi.org/10.17504/protocols.io.x23fqgn 2021-03-29 03:15:01
U Mass - Glucose Tolerance Test with insulin secretion
 
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Jason Kim 10.17504/protocols.io.xxbfpin Mouse Metabolic Phenotyping Centers Summary:Glucose tolerance test with insulin secretion measures systemic clearance of glucose and systemic appearance of insulin following an intraperitoneal bolus injection of 20% dextrose. This experiment measures insulin sensitivity and insulin secretion (pancreatic -cell function) in awake mice assuming that there are no alterations in systemic insulin clearance. Insulin sensitivity and pancreatic -cell function are altered in obese mice. RRID:AB_2792981 University of Massachusetts https://mmpc.org/shared/document.aspx?id=142&docType=Protocol 1 2019 Jason Kim 2019. U Mass - Glucose Tolerance Test with insulin secretion. protocols.io dx.doi.org/10.17504/protocols.io.xxbfpin 2021-03-29 03:13:07
Yale - HDL Cholesterol
 
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John Stack, Gary Cline 10.17504/protocols.io.y3ufynw Mouse Metabolic Phenotyping Centers Procedure used to determine the concentration of HDL cholesterol in blood, serum, and plasma. HDL Cholesterol is determined in a two-step procedure. First chylomicrons, VLDL, and LDL are selectively reacted with cholesterol esterase and eliminated from the reaction. In the second step, the remaining HDL-cholesterol is assayed as described for total cholesterol. Yale University, Yale University https://mmpc.org/shared/document.aspx?id=210&docType=Protocol 1 2019 John Stack, Gary Cline 2019. Yale - HDL Cholesterol. protocols.io dx.doi.org/10.17504/protocols.io.y3ufynw 2021-03-29 03:13:09
U Mass - Non-esterified fatty acids
 
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Jason Kim 10.17504/protocols.io.x3hfqj6 Mouse Metabolic Phenotyping Centers This experiment measures serum and plasma concentrations of non-esterified fatty acids using a 96-well kit. The experiment involves a coupled reaction to measure non-esterified fatty acids (NEFA) which ultimately forms a purple product that absorbs light at 550nm. This allows the concentration of NEFA to be determined from the optical density measured at 540~550nm. Serum fatty acids levels reflect systemic lipid metabolism, lipid digestion/absorption, and lipid clearance. Serum fatty acids levels are altered in obesity, insulin resistance, and type 2 diabetes. University of Massachusetts http://mmpc.org/shared/document.aspx?id=168&docType=Protocol 1 2019 Jason Kim 2019. U Mass - Non-esterified fatty acids. protocols.io dx.doi.org/10.17504/protocols.io.x3hfqj6 2021-03-29 03:13:07
U Mass - Total protein
 
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Jason Kim 10.17504/protocols.io.x5gfq3w Mouse Metabolic Phenotyping Centers This experiment involves a spectrophotometric measurement using Roche Cobas Clinical Chemistry Analyzer. Serum levels of total protein reflect systemic protein metabolism. University of Massachusetts https://mmpc.org/shared/document.aspx?id=186&docType=Protocol 1 2019 Jason Kim 2019. U Mass - Total protein. protocols.io dx.doi.org/10.17504/protocols.io.x5gfq3w 2021-03-29 03:13:08
U Mass - Body composition (organs)
 
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Jason Kim 10.17504/protocols.io.xsnfnde Mouse Metabolic Phenotyping Centers The EchoMRI 3-in-1 uses ¹H- magnetic resonance spectroscopy to noninvasively measure fat mass, lean mass, and water mass in individual organs. Fat, lean, and water mass composition in specific organs is altered in obesity. University of Massachusetts https://mmpc.org/shared/document.aspx?id=148&docType=Protocol 1 2019 Jason Kim 2019. U Mass - Body composition (organs). protocols.io dx.doi.org/10.17504/protocols.io.xsnfnde 2021-03-29 03:13:10
U Mass - Triglyceride
 
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Jason Kim 10.17504/protocols.io.x5pfq5n Mouse Metabolic Phenotyping Centers, Metabolomics Protocols & Workflows This experiment involves a spectrophotometric measurement using Roche Cobas Clinical Chemistry Analyzer. Triglyceride levels may be measured in serum, plasma, and tissue samples. Serum and tissue triglyceride levels are affected by alterations in systemic lipid metabolism, lipid digestion/absorption, and lipid clearance. Serum and tissue triglyceride levels are altered in obesity, insulin resistance, type 2 diabetes, alcoholic steatohepatitis, non-alcoholic fatty liver disease, and non-alcoholic steatohepatitis. University of Massachusetts https://mmpc.org/shared/document.aspx?id=167&docType=Protocol 1 2019 Jason Kim 2019. U Mass - Triglyceride. protocols.io dx.doi.org/10.17504/protocols.io.x5pfq5n 2021-03-29 03:13:09
UC Davis - Basic Adipocyte Culture Protocol
 
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K.C. Kent Lloyd 10.17504/protocols.io.7bvhin6 Mouse Metabolic Phenotyping Centers Basic Adipocyte Culture Protocol University of California, Davis https://mmpc.org/shared/document.aspx?id=82&docType=Protocol 2 2019 K.C. Kent Lloyd 2019. UC Davis - Basic Adipocyte Culture Protocol. protocols.io dx.doi.org/10.17504/protocols.io.7bvhin6 2021-03-29 03:13:53
U Michigan - Hind Paw Withdrawal for Rodents
 
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Eva Feldman 10.17504/protocols.io.x67frhn Mouse Metabolic Phenotyping Centers This protocol measures and quantifies the pain in rodent hind paw. These measurements can be used to evaluate the pathology present in the nervous system. University of Michigan - Ann Arbor https://mmpc.org/shared/document.aspx?id=318&docType=Protocol 1 2019 Eva Feldman 2019. U Michigan - Hind Paw Withdrawal for Rodents. protocols.io dx.doi.org/10.17504/protocols.io.x67frhn 2021-03-29 03:13:57
Case - Intraperitoneal Glucose Tolerance Test
 
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Henri Brunengraber 10.17504/protocols.io.ydtfs6n Mouse Metabolic Phenotyping Centers This is the standard protocol for most routine glucose tolerance testing. It is performed on awake mice, fasted for 18 hr (overnight) or 6 hours. Case Western Reserve University https://mmpc.org/shared/document.aspx?id=271&docType=Protocol 1 2019 Henri Brunengraber 2019. Case - Intraperitoneal Glucose Tolerance Test. protocols.io dx.doi.org/10.17504/protocols.io.ydtfs6n 2021-03-29 03:14:16
UC Davis - Glutathione Reductase
 
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Peter Havel 10.17504/protocols.io.68ehhte Mouse Metabolic Phenotyping Centers Summary: Glutathione reductase (GR, EC 1.6.4.2) is a flavoprotein that catalyzes the NADPH- dependent reduction of oxidized glutathione (GSSG) to glutathione (GSH). This enzyme is essential for the GSH redox cycle which maintains adequate levels of reduced cellular GSH. A high GSH/GSSG ratio is essential for protection against oxidative stress. The Cayman Chemical Glutathione Reductase Assay Kit measures GR activity by measuring the rate of NADPH oxidation. The oxidation of NADPH to NADP+ is accompanied by a decrease in absorbance at 340 nm. Since GR is present at rate limiting concentrations, the rate of decrease in the A340 is directly proportional to the GR activity in the sample. The Cayman GR Assay Kit is provided in a convenient 96 well plate format and can be used to measure GR activity inplasma, erythrocyte lysates, tissue homogenates, and cell lysates. University of California, Davis https://mmpc.org/shared/document.aspx?id=127&docType=Protocol 2 2019 Peter Havel 2019. UC Davis - Glutathione Reductase. protocols.io dx.doi.org/10.17504/protocols.io.68ehhte 2021-03-29 03:14:11
Vandy - Mouse Blood Pressure Telemetry
 
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Lin Zhong, Chee Lim 10.17504/protocols.io.6xnhfme Mouse Metabolic Phenotyping Centers Summary:Imlantable radiotelemetry devices allow for long-term continous monitoring of blood pressure and heart rate in conscious freely moving caged animals. This protocol describes the surgical implantation of a radio transmitter device in the mouse. Vanderbilt University, Vanderbilt University https://mmpc.org/shared/document.aspx?id=229&docType=Protocol 2 2019 Lin Zhong, Chee Lim 2019. Vandy - Mouse Blood Pressure Telemetry. protocols.io dx.doi.org/10.17504/protocols.io.6xnhfme 2021-03-29 03:15:17
UC Davis - Intraperitoneal Glucose Tolerance Test
 
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Fawaz G. Haj 10.17504/protocols.io.yp4fvqw Mouse Metabolic Phenotyping Centers Summary: An intraperitoneal Glucose tolerance test or ipGTT is designed to determine clearance of an intraperitoneally injected glucose load from the body. Animals are fasted for approximately 16 hours, a solution of glucose is administered by intraperitoneal (IP) injection and blood glucose is measured at different time points during 2 hours post-injection. University of California, Davis https://mmpc.org/shared/document.aspx?id=85&docType=Protocol 1 2019 Fawaz G. Haj 2019. UC Davis - Intraperitoneal Glucose Tolerance Test. protocols.io dx.doi.org/10.17504/protocols.io.yp4fvqw 2021-03-29 03:15:30
U Michigan - Illumina 16S rRNA gene sequencing using DNA
 
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Vincent Young 10.17504/protocols.io.7bzhip6 Mouse Metabolic Phenotyping Centers Summary: This protocol is for the submission of DNA to generate libraries for 16S rRNA sequencing, which can be used for bacterial community analysis and detect variations in the microbiota under differing conditions.DNA is aliquoted into 96 well plates, which are properly packaged and shipped to the MMPC and libraries are prepared for community analysis as previously described by Seekatz et al. (1). Briefly, barcoded dual-index primers specific to the V4 region of the 16S rRNA gene amplify the DNA (2). PCR reactions are composed of 5 μL of 4 μM equimolar primer set, 0.15 μL of AccuPrime Taq DNA High Fidelity Polymerase, 2 μL of 10x AccuPrime PCR Buffer II (Thermo Fisher Scientific, catalog no. 12346094), 11.85 μL of PCR-grade water, and 1 μL of DNA template. The PCR conditions used consisted of 2 min at 95°C, followed by 30 cycles of 95°C for 20 s, 55°C for 15 s, and 72°C for 5 min, followed by 72°C for 10 min. Each PCR reaction is normalized using the SequalPrep Normalization Plate Kit (Thermo Fisher Scientific, catalog no. A1051001). The normalized reactions are pooled and quantified using the Kapa Biosystems Library qPCR MasterMix (ROX Low) Quantification kit for Illumina platforms (catalog no. KK4873). The Agilent Bioanalyzer is used to confirm the size of the amplicon library (~399 bp) using a high-sensitive DNA analysis kit (catalog no. 5067-4626). Pooled amplicon library is then sequenced on the Illumina MiSeq platform using the 500 cycle MiSeq V2 Reagent kit (catalog no. MS-102-2003) according to the manufacturer's instructions with modifications of the primer set with custom read 1/read 2 and index primers added to the reagent cartridge. The“Preparing Libraries for Sequencing on the MiSeq” (part 15039740, Rev. D) protocol was used to prepare libraries with a final load concentration of 5.5 pM, spiked with 15% PhiX to create diversity within the run. FASTQ files are distributed to the client when the 2 x 250 bp sequencing completes.References:1. Seekatz AM, Theriot CM, Molloy CT, Wozniak KL, Bergin IL, Young VB. 2015. Fecal Microbiota Transplantation Eliminates Clostridium difficile in a Murine Model of Relapsing Disease. Infect Immun 83:3838-3846. 10.1128/IAI.00459-15. 2. Kozich JJ, Westcott SL, Baxter NT, Highlander SK, Schloss PD. 2013. Development of a dual- index sequencing strategy and curation pipeline for analyzing amplicon sequence data on the MiSeq Illumina sequencing platform. Appl Environ Microbiol 79:5112–5120. 10.1128/AEM.01043-13. University of Michigan - Ann Arbor https://mmpc.org/shared/document.aspx?id=305&docType=Protocol 2 2019 Vincent Young 2019. U Michigan - Illumina 16S rRNA gene sequencing using DNA. protocols.io dx.doi.org/10.17504/protocols.io.7bzhip6 2021-03-29 03:15:28
U Cinn - NEFA Concentration
 
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Patrick Tso, Dana Lee 10.17504/protocols.io.xmnfk5e Mouse Metabolic Phenotyping Centers Quantitative determinations of non-esterified fatty acids in plasma/serum/lymph will be made using the NEFA-HR enzymatic colorimetric method assay. University of Cincinnati, University of Cincinnati https://mmpc.org/shared/document.aspx?id=196&docType=Protocol 1 2019 Patrick Tso, Dana Lee 2019. U Cinn - NEFA Concentration. protocols.io dx.doi.org/10.17504/protocols.io.xmnfk5e 2021-03-29 03:12:30
UC Davis - Blood Pressure by Tail Cuff
 
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Jennifer Rutkowsky 10.17504/protocols.io.yetften Mouse Metabolic Phenotyping Centers This technique has been used routinely for the non-invasive measurement of blood pressure in rats, and more recently in mice. The technique provides a good estimate of actual systolic pressure. Hypertension major risk factor for stroke myocardial infarction, heart failure, aneurysm of the arteries, peripheral artery disease and is a cause of chronic kidney disease and erectile dysfunction (ED)Modified from: UC Davis MBP SOP-CODA Blood Pressure, Todd Tolentino 11/02/2012 University of California, Davis https://mmpc.org/shared/document.aspx?id=110&docType=Protocol 1 2019 Jennifer Rutkowsky 2019. UC Davis - Blood Pressure by Tail Cuff. protocols.io dx.doi.org/10.17504/protocols.io.yetften 2021-03-29 03:12:32
U Mass - Chronic/acute phloridzin treatment
 
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Jason Kim 10.17504/protocols.io.xujfnun Mouse Metabolic Phenotyping Centers Summary:Phloridzin is a potent inhibitor of renal glucose reabsorption and may be used to lower serum glucose levels. Phloridzin may be administered chronically to induce glycosuria and lower hyperglycemia in diabetic mice. Phloridzin may be administered acutely to reduce serum glucose levels for experiments in diabetic mice. University of Massachusetts https://mmpc.org/shared/document.aspx?id=154&docType=Protocol 1 2019 Jason Kim 2019. U Mass - Chronic/acute phloridzin treatment. protocols.io dx.doi.org/10.17504/protocols.io.xujfnun 2021-03-29 03:12:30

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