Protocols   Select Another Resource Report Type

Authors: John Stack, Gary Cline
Group: Mouse Metabolic Phenotyping Centers
Summary: Procedure used to determine the concentration of calcium in blood, serum, and plasma. Calcium is measured as the complex with arsenazo III and monitored at 600nm.

Proper citation: John Stack, Gary Cline 2019. Yale - Blood or Urine Calcium. protocols.io https://dx.doi.org/10.17504/protocols.io.y3nfyme Copy   

•   Source:

Authors: Henri Brunengraber
Group: Mouse Metabolic Phenotyping Centers
Summary: Mouse Insulin ELISA kit is used for the non radioactive quantification of insulin in mouse plasma.

Proper citation: Henri Brunengraber 2019. Case - Plasma Insulin Measurement by ELISA. protocols.io https://dx.doi.org/10.17504/protocols.io.yesftee Copy   

•   Source:

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   

•   Source:

Authors: Jeff Hodgin
Group: Mouse Metabolic Phenotyping Centers
Summary: This is the protocol for measuring Glomerular Filtration Rate (GFR) with minipump FITC-inulin clearance in mice. In brief, Alzet micro-osmotic pumps (minipumps, Model 1007D) are filled with 3% FITC-inulin solution. These pumps release FITC-inulin at a rate of 0.5 μl/hr at least seven days. Mouse is temporarily anesthetized and then two mini-pumps are implanted in mouse peritoneal cavity through a tiny midline incision. After surgery, 24-hour urine is collected with metabolic cage and 100 μl of blood from saphenous vein is obtained at the same time. The concentrations of FITC-inulin in urine and blood samples are measured by flurometer and GFR is calculated by the concentration of FITC-inulin in 24-hour-urine and the concentration of FITC-inulin in plasma.

Proper citation: Jeff Hodgin 2019. U Michigan - Glomerular Filtration Rate Determination with Minipump Inulin Clearance. protocols.io https://dx.doi.org/10.17504/protocols.io.x63frgn Copy   

•   Source:

Authors: Peter Havel
Group: Mouse Metabolic Phenotyping Centers
Summary: Cholesterol esters are enzymatically hydrolysed by cholesterol esterase to cholesterol and free fatty acids. Free cholesterol, including that originally present, is then oxidized by cholesterol oxidase to cholest-4-en-3one and hydrogen peroxide. The hydrogen peroxide combines with HBA and 4-aminoantipyrine to form a chromophore (quinoneimine dye) which may be quantitated at 500-550nm.

Proper citation: Peter Havel 2019. UC Davis - Total Cholesterol (TC) Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.ygeftte Copy   

•   Source:

Authors: Fawaz G. Haj
Group: Mouse Metabolic Phenotyping Centers
Summary: Summary: This test is designated to determine if rodents exhibit activation of leptin signaling pathway through evaluation of the phosphorylation state of JAK2 and STAT3 proteins.

Proper citation: Fawaz G. Haj 2019. UC Davis - Leptin Signaling pathway. protocols.io https://dx.doi.org/10.17504/protocols.io.ybqfsmw Copy   

•   Source:

Authors: Gary Cline, John Stack
Group: Mouse Metabolic Phenotyping Centers
Summary: Procedure used to measure the activity of Aspartate Amino Transferase (AST). AST activity is measured by the enzymatically coupled reactions of AST (to form oxaloacetate from aspartate and α-ketoglutarate) and malate dehydrogenase (conversion of oxaloacetate to malate with oxdiation of NADH to NAD). The rate of NAD formation is monitored by the change in absorbance at 340 nm.

Proper citation: Gary Cline, John Stack 2019. Yale - Aspartate Amino Transferase. protocols.io https://dx.doi.org/10.17504/protocols.io.yz9fx96 Copy   

•   Source:

Authors: John Stack, Gary Cline
Group: Mouse Metabolic Phenotyping Centers
Summary: Procedure used to determine the total concentration of cholesterol present in blood, serum, and plasma. Cholesterol esters are hydrolysed by cholesterol esterase. Cholesterol is then oxidized by cholesterol oxidase with formation of hydrogen peroxide. Peroxidase uses the hydrogen peroxide, phenol, and 4-aminotipyrine to form a quinoneimine dye which is measured at 500 nm.

Proper citation: John Stack, Gary Cline 2019. Yale - Total Cholesterol. protocols.io https://dx.doi.org/10.17504/protocols.io.y3rfym6 Copy   

•   Source:

Authors: Oliver Fiehn
Group: Mouse Metabolic Phenotyping Centers, Metabolomics Protocols & Workflows
Summary: Description of standard setting for analysis of derivatized samples ready for injection into a Leco Pegasus III or Pegasus IV gas chromatograph-time of flight mass spectrometer.References: Fiehn O, Wohlgemuth G, Scholz M, Kind T, Lee DY, Lu Y, Moon S, Nikolau BJ (2008) Quality control for plant metabolomics: Reporting MSI-compliant studies. Plant J. 53, 691-704

Proper citation: Oliver Fiehn 2019. UC Davis - Metabolomics: Primary metabolites by GC-TOF MS. protocols.io https://dx.doi.org/10.17504/protocols.io.64nhgve Copy   

•   Source:

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   

•   Source:

Authors: John Rutledge
Group: Mouse Metabolic Phenotyping Centers
Summary: Dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) is used to assess blood-brain barrier permeability in vivo in rodent models. Post-processing image analysis is done using our in-house matlab script with patlak linearized regression mathematical modeling.

Proper citation: John Rutledge 2019. UC Davis - Dynamic contrast enhanced magnetic resonance imaging. protocols.io https://dx.doi.org/10.17504/protocols.io.yh9ft96 Copy   

•   Source:

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   

•   Source:

Authors: Vance L. Albaugh
Group: Mouse Metabolic Phenotyping Centers
Summary: This is the protocol for conducting the bile diversion procedure in the mouse. In this procedure the common bile duct is ligated proximal to the confluence of the common bile and the pancreatic ducts and an anastomosis is created at a prespecified point along the gastrointestinal tract to re-establish drainage of the biliary tree.

Proper citation: Vance L. Albaugh 2019. Vandy – Bile Diversion in Mice. protocols.io https://dx.doi.org/10.17504/protocols.io.zdcf22w Copy   

•   Source:

Authors: Patrick Tso, Dana Lee
Group: Mouse Metabolic Phenotyping Centers
Summary: Dietary fat containing 5% sucrose polybehenate, a non-absorbable food additive, is fed in a semi-synthetic diet containing absorbable fat to rodents. The animals eat the diet ad lib for 3 days. Two or more fecal pellets are collected on days 3 and 4. Fat absorption is calculated from the ratios of behenic acid to other fatty acids in the diet and in the feces as analyzed by gas chromatography of fatty acid methyl esters.

Proper citation: Patrick Tso, Dana Lee 2019. U Cinn - Non-invasive Measurement of Intestinal Fat Absorption. protocols.io https://dx.doi.org/10.17504/protocols.io.xmtfk6n Copy   

•   Source:

Authors: Peter Havel
Group: Mouse Metabolic Phenotyping Centers
Summary: Summary: Alanine transaminase (ALT) also known as alanine aminotransferase or (sGPT) is a metabolic enzyme expressed primarily in the liver. Elevation of ALT levels is an indication of liver damage and has been associated with liver injury. ALT levels are monitored routinely in patients with liver diseases. ALT is also a very useful tool for preclinical investigation of experimental drug formulations and ALT levels are commonly used to monitor and attenuate the hepatotoxic effects of experimental drugs in rodents.

Proper citation: Peter Havel 2019. UC Davis - Alanine transaminase. protocols.io https://dx.doi.org/10.17504/protocols.io.ybtfsnn Copy   

•   Source:

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   

•   Source:

Authors: John Stack, Gary Cline
Group: Mouse Metabolic Phenotyping Centers
Summary: Procedure used to measure glucose concentrations. Glucose is measured by the enzymatically coupled reactions of hexokinase and glucose-6-P dehydrogenase. The rate of NADH formation is monitored by the change in absorbance at 340 nm.

Proper citation: John Stack, Gary Cline 2019. Yale - Blood Glucose. protocols.io https://dx.doi.org/10.17504/protocols.io.y2kfycw Copy   

•   Source:

Authors: Peter Havel
Group: Mouse Metabolic Phenotyping Centers
Summary: Lipoprotein profiling and Lipoprotein size will be provided for CM, VLDL, LDL and HDL as well as 20 additional subclasses of lipoprotein. Testing requires as little as10µL for full analytical testing.

Proper citation: Peter Havel 2019. UC Davis - Lipoprotein profiling and partical size. protocols.io https://dx.doi.org/10.17504/protocols.io.yrifv4e Copy   

•   Source:

Authors: Lin Zhong, Jeffrey Rottman, Chee Lim
Group: Mouse Metabolic Phenotyping Centers
Summary: Summary:The most common cause of cardiovascular mortality in man is the outcome from myocardial ischemic injury. Accordingly, it is necesarry to study the corresponding process of heart injury recovery in many mouse models relevant to human cardiovascular disease. This protocol describes the surgical induction of myocardial injury via transient occlusion of a coronary artery followed by reperfusion (ischemia-reperfusion injury).

Proper citation: Lin Zhong, Jeffrey Rottman, Chee Lim 2019. Vandy - Myocardial Ischemia Reperfusion. protocols.io https://dx.doi.org/10.17504/protocols.io.6xfhfjn Copy   

•   Source:

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   

•   Source: