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| Name | Authors | DOI | Group |
Summary |
Associated Publications |
RRIDs used | ||||||
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UC Davis - LDL Protocol Resource Report Resource Website |
Peter Havel | 10.17504/protocols.io.yrcfv2w | Mouse Metabolic Phenotyping Centers | LDL and VLDL are separated from HDL using a precipitation reagent. Then the HDL fraction is measured for either TC or TG using the same reagents for total cholesterol or triglyceride. | University of California, Davis | https://mmpc.org/shared/document.aspx?id=94&docType=Protocol | 1 | 2019 | Peter Havel 2019. UC Davis - LDL Protocol. protocols.io dx.doi.org/10.17504/protocols.io.yrcfv2w | 2021-03-29 03:12:41 | ||
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UC Davis - Metabolomics: Sample preparation for GCTOF analysis Resource Report Resource Website |
Oliver Fiehn | 10.17504/protocols.io.ytjfwkn | Mouse Metabolic Phenotyping Centers, Metabolomics Protocols & Workflows | This SOP describes sample extraction and sample preparation for primary metabolism profiling by gas chromatography / time of flight mass spectrometry (GCTOF)References: Fiehn O, Kind T (2006) Metabolite profiling in blood plasma. In: Metabolomics: Methods and Protocols. Weckwerth W (ed.), Humana Press, Totowa NJ | University of California, Davis | https://mmpc.org/shared/document.aspx?id=121&docType=Protocol | 1 | 2019 | Oliver Fiehn 2019. UC Davis - Metabolomics: Sample preparation for GCTOF analysis. protocols.io dx.doi.org/10.17504/protocols.io.ytjfwkn | 2021-03-29 03:12:43 | ||
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Yale - Triglycerides Resource Report Resource Website |
John Stack, Gary Cline | 10.17504/protocols.io.y4bfysn | Mouse Metabolic Phenotyping Centers | Procedure used to determine the concentration of triglycerides in blood, serum, and plasma. Triglycerides are determined by coupling lipase, glucokinase, glycerol phosphate oxidase, and peroxidase to form a quinonemine dye which is measured at 500 nm. | Yale University, Yale University | https://mmpc.org/shared/document.aspx?id=213&docType=Protocol | 1 | 2019 | John Stack, Gary Cline 2019. Yale - Triglycerides. protocols.io dx.doi.org/10.17504/protocols.io.y4bfysn | 2021-03-29 03:12:43 | ||
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U Michigan - Western Blot Resource Report Resource Website |
Jeff Hodgin | 10.17504/protocols.io.56hg9b6 | Mouse Metabolic Phenotyping Centers | Immunoblotting (also called Western Blotting) is a technique to separate and identify individual proteins in a protein mixture (e.g. a cell lysate). In Immunoblotting (Western blotting) the protein mixture is applied to a gel electrophoresis (SDS-PAGE gel electrophoresis) to sort the proteins by its molecular weight size in individual protein bands. The separated protein bands are then transferred to a nitrocellulose transfer membrane. The proteins adhere to the membrane in the same pattern as they have been separated due to interactions of charges. To visualize the protein of interest the membrane is commonly first probed using a primary protein- specific antibody followed by a labeled secondary antibody. The specificity of the antibody-antigen interaction enables a target protein to be identified in the midst of a complex protein mixture. The bound antibodies are then detected by western blotting substrate (e.g. Pierce ECL Western Blotting Substrate), and then visualized by using adeveloping film (HyBlot CL Autoradiography Film) in the darkroom. The thickness of the band corresponds to the amount of protein present. | University of Michigan - Ann Arbor | https://mmpc.org/shared/document.aspx?id=316&docType=Protocol | 2 | 2019 | Jeff Hodgin 2019. U Michigan - Western Blot. protocols.io dx.doi.org/10.17504/protocols.io.56hg9b6 | 2021-03-29 03:12:45 | ||
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U Mass - Alkaline Phosphatase Resource Report Resource Website |
Jason Kim | 10.17504/protocols.io.xptfmnn | Mouse Metabolic Phenotyping Centers | This experiment involves a spectrophotometric measurement using Roche Cobas Clinical Chemistry Analyzer. Serum levels of alkaline phosphatase reflect liver function. | University of Massachusetts | https://mmpc.org/shared/document.aspx?id=177&docType=Protocol | 1 | 2019 | Jason Kim 2019. U Mass - Alkaline Phosphatase. protocols.io dx.doi.org/10.17504/protocols.io.xptfmnn | 2021-03-29 03:12:46 | ||
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UC Davis - Protein Carbonyl Resource Report Resource Website |
Peter Havel | 10.17504/protocols.io.63hhgj6 | Mouse Metabolic Phenotyping Centers | Summary: Cayman Chemical's Protein Carbonyl Colorimetric Assay Kit is a convenient colorimetric assay for the measurement of oxidized proteins. Protein samples are derivatized by making use of the reaction between 2,4-dinitrophenylhydrazine (DNPH) and protein carbonyls. Formation of a Schiff base produces the corresponding hydrazone which can be analyzed spectrophotometrically at 360-385 nm. This assay can be used to measure oxidized protein in plasma, serum, cell lysates, and tissue homogenates. | University of California, Davis | https://mmpc.org/shared/document.aspx?id=123&docType=Protocol | 2 | 2019 | Peter Havel 2019. UC Davis - Protein Carbonyl. protocols.io dx.doi.org/10.17504/protocols.io.63hhgj6 | 2021-03-29 03:12:51 | ||
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Vandy - Hyperinsulinemic-euglycemic Clamp Resource Report Resource Website |
Li Kang | 10.17504/protocols.io.yyafxse | Mouse Metabolic Phenotyping Centers | Summary:Mice with catheters implanted in the jugular vein (infusions) and carotid artery (sampling) are used for this procedure (V3002). The hyperinsulinemic, euglycemic clamp or “insulin clamp” has been used in a variety of species to assess insulin action. In an insulin clamp, the rate of glucose infused to maintain euglycemia is an index of whole body insulin sensitivity. Isotopes can be used during an insulin clamp to distinguish between insulin’s effects on endogenous glucose production and glucose utilization, or to examine insulin’s effects on specific tissues and metabolic pathways. The insulin infusion rate used in a study depends on a number of factors such as whether the experiment requires a sensitive analysis of endogenous glucose production, or whether the mouse is known to be unusually insulin resistant (e.g. ob/ob). Finally the insulin clamp technique, especially when isotopes are involved, is most quantitative under steady state conditions. Therefore, clamps should be of a duration (~2 h) that is sufficient for the actions of insulin to obtain steady state. The section that follows describes, a) the technical procedures used on the day of a clamp experiment; b) preparation of insulin and tracer infusates; and c) analyses of plasma and tissue radioactivity. Familiarity with these procedures combined with laboratory demonstration provides the investigator with the necessary background to clamp the conscious mouse. | Vanderbilt University | https://mmpc.org/shared/document.aspx?id=239&docType=Protocol | 1 | 2019 | Li Kang 2019. Vandy - Hyperinsulinemic-euglycemic Clamp. protocols.io dx.doi.org/10.17504/protocols.io.yyafxse | 2021-03-29 03:13:03 | ||
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UC Davis - Radial Arm Water Maze Resource Report Resource Website |
Mari Golub | 10.17504/protocols.io.yv2fw8e | Mouse Metabolic Phenotyping Centers | The radial arm water maze (RAWM) measures spatial learning by combining both a water maze and radial arm maze (RAM). This test measures both working and reference memory through a reward system, in this case escape from water. This method is advantageous to other RAM which use food as a reward in that it may be utilized with mice on specialized diets. Reference: Behavioral consequences of ovarian atrophy and estrogen replacement in the APPsswe mouse. Golub et al. Neurobiol Aging (2008) 29(10): 1512-1523. | University of California, Davis | https://mmpc.org/shared/document.aspx?id=277&docType=Protocol | 1 | 2019 | Mari Golub 2019. UC Davis - Radial Arm Water Maze. protocols.io dx.doi.org/10.17504/protocols.io.yv2fw8e | 2021-03-29 03:12:59 | ||
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U Mass - Coronary Artery Ligation Resource Report Resource Website |
Mark Kelly LAT, Timothy P. Fitzgibbons | 10.17504/protocols.io.56kg9cw | Mouse Metabolic Phenotyping Centers | Summary: This is a mouse model of acute myocardial infarction. This protocol is suitable for studying mouse models thought to be relevant to coronary ischemia or the development of congestive heart failure. | University of Massachusetts Medical School, University of Massachusetts Medical School | https://mmpc.org/shared/document.aspx?id=325&docType=Protocol | 2 | 2019 | Mark Kelly LAT, Timothy P. Fitzgibbons 2019. U Mass - Coronary Artery Ligation. protocols.io dx.doi.org/10.17504/protocols.io.56kg9cw | 2021-03-29 03:13:01 | ||
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UC Davis - Aspartate Aminotransferase Resource Report Resource Website |
Peter Havel | 10.17504/protocols.io.ybvfsn6 | Mouse Metabolic Phenotyping Centers | Summary: Aspartate transaminase (AST) also known as aspartate aminotransferase or (sGOT) is a metabolic enzyme expressed primarily in the liver. Elevation of AST levels is an indication of liver damage and has been associated with liver injury. AST levels are monitored routinely in patients with liver diseases. AST is also a very useful tool for preclinical investigation of experimental drug formulations and AST levels are commonly used to monitor and attenuate the hepatotoxic effects of experimental drugs in rodents. | University of California, Davis Metabolism and Endocrinology Core | https://mmpc.org/shared/document.aspx?id=290&docType=Protocol | 1 | 2019 | Peter Havel 2019. UC Davis - Aspartate Aminotransferase. protocols.io dx.doi.org/10.17504/protocols.io.ybvfsn6 | 2021-03-29 03:13:02 | ||
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UC Davis - Acquisition/Acclimation/Quarantine Resource Report Resource Website |
K.C. Kent Lloyd | 10.17504/protocols.io.xfifjke | Mouse Metabolic Phenotyping Centers | SummaryThe purpose of this standard operating procedure (SOP) is to provide steps for the acquisition, acclimation and quarantine of animals obtained from other institutions or laboratories and introduced to the MBP Vivarium with minimal risk to the current mouse colony population. This procedure applies to the import/export staff, veterinary staff and any principal investigator requesting animals. | University of California, Davis | https://www.mmpc.org/shared/document.aspx?id=117&docType=Protocol | 1 | 2019 | K.C. Kent Lloyd 2019. UC Davis - Acquisition/Acclimation/Quarantine. protocols.io dx.doi.org/10.17504/protocols.io.xfifjke | 2021-03-29 03:13:16 | ||
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U Mass - Ammonia Resource Report Resource Website |
Jason Kim | 10.17504/protocols.io.xq3fmyn | Mouse Metabolic Phenotyping Centers | Summary:This experiment involves a spectrophotometric measurement using Roche Cobas Clinical Chemistry Analyzer. | University of Massachusetts | https://mmpc.org/shared/document.aspx?id=172&docType=Protocol | 1 | 2019 | Jason Kim 2019. U Mass - Ammonia. protocols.io dx.doi.org/10.17504/protocols.io.xq3fmyn | 2021-03-29 03:13:25 | ||
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U Mass - Amylase Resource Report Resource Website |
Jason Kim | 10.17504/protocols.io.xq4fmyw | Mouse Metabolic Phenotyping Centers | This experiment involves a spectrophotometric measurement using Roche Cobas Clinical Chemistry Analyzer. | University of Massachusetts | https://mmpc.org/shared/document.aspx?id=175&docType=Protocol | 1 | 2019 | Jason Kim 2019. U Mass - Amylase. protocols.io dx.doi.org/10.17504/protocols.io.xq4fmyw | 2021-03-29 03:13:28 | ||
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UC Davis - Gut Microbiome Analysis (Illumina-20K reads, 2X300bp) Resource Report Resource Website |
Trina Knotts | 10.17504/protocols.io.ynpfvdn | Mouse Metabolic Phenotyping Centers, Microbiome Centers | The work of Gordon and colleagues (i.e., Nature. 2006 Dec 21;444(7122):1027-31) has shown that obesity can result in marked shifts in the gut microbiome in mice and other models including humans. While the role of the microbiome remains to be fully elucidated, the gut microbiota can no longer be ignored as a potentially important factor when assessing metabolic phenotype. This service involves 16S gene variable region (V3-V4 using primer pair 515F-806R) pair end sequencing (2x300bp) by Illumina (20,000 avg seq read depth) of feces, cecal, or other GI contents. Sequences can be processed through a bioinformatics pipeline (Qiime) to taxonomically classify them and to assess alpha and beta diversity of the community. In addition, Principal Components Analysis (PCA) or partial least squares- discriminant analysis (PLS-DA) can leverage variances in the relative microbial abundances to better understand how specific microbes contribute to separation by group. Correlational analyses can identify which variables of host metadata associate with specific microbes. The Core’s gut microbiome assay will employ this approach to uncover unique microbiota fingerprints in test mice. One caveat is that with fecal samples, patterns are only a surrogate for actual gut microbiota patterns, and may not exactly reflect the intestinal populations. | University of California, Davis | https://mmpc.org/shared/document.aspx?id=288&docType=Protocol | 1 | 2019 | Trina Knotts 2019. UC Davis - Gut Microbiome Analysis (Illumina-20K reads, 2X300bp). protocols.io dx.doi.org/10.17504/protocols.io.ynpfvdn | 2021-03-29 03:13:36 | ||
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U Michigan - Mesangial matrix evalutaion Resource Report Resource Website |
Jeff Hodgin, Jharna Saha | 10.17504/protocols.io.562g9ge | Mouse Metabolic Phenotyping Centers | Periodic acid-Schiff (PAS) is a staining method used to detect glycogen on formalin-fixed, paraffin-embedded kidney tissue sections. PAS staining highlights basement membranes and is frequently used to diagnose glomerular mesangial matrix expansion. | University of Michigan - Ann Arbor, Previous Brosius lab member | https://mmpc.org/shared/document.aspx?id=312&docType=Protocol | 2 | 2019 | Jeff Hodgin, Jharna Saha 2019. U Michigan - Mesangial matrix evalutaion. protocols.io dx.doi.org/10.17504/protocols.io.562g9ge | 2021-03-29 03:13:43 | ||
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Yale - LDL Cholesterol Resource Report Resource Website |
John Stack, Gary Cline | 10.17504/protocols.io.y3vfyn6 | Mouse Metabolic Phenotyping Centers | Procedure used to determine the concentration of LDL cholesterol in blood, serum, and plasma. LDL-cholesterol is determined by an elimination method to selectively remove HDL, VLDL, and IDL subfractions prior to assay of LDL-cholesterol as described for total cholesterol. | Yale University, Yale University | https://mmpc.org/shared/document.aspx?id=211&docType=Protocol | 1 | 2019 | John Stack, Gary Cline 2019. Yale - LDL Cholesterol. protocols.io dx.doi.org/10.17504/protocols.io.y3vfyn6 | 2021-03-29 03:13:42 | ||
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UC Davis - Mouse Gross Necropsy with Histology Resource Report Resource Website |
Stephen M. Griffey | 10.17504/protocols.io.yusfwwe | Mouse Metabolic Phenotyping Centers | Summary: Gross necropsy with documentation of changes present. Collection and histologic processing (paraffin blocks/H&E slides) and interpretation of kidneys, liver, spleen, pancreas, heart, lungs, esophagus, trachea, thymus, mesenteric lymph nodes, GI Tract, cerebrum, cerebellum, urinary bladder, and reproductive tract and additional tissues with gross changes. This includes photodocumentation of significant findings. This assay can be customized to include other target organs based on findings from other MMPC assays. This procedure is to be followed by trained Comparative Pathology Laboratory (CPL) staff responsible for performing necrospy diagnostics procedures at CPL. | University of California, Davis | https://mmpc.org/shared/document.aspx?id=132&docType=Protocol | 1 | 2019 | Stephen M. Griffey 2019. UC Davis - Mouse Gross Necropsy with Histology. protocols.io dx.doi.org/10.17504/protocols.io.yusfwwe | 2021-03-29 03:13:44 | ||
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U Mass - Alanine Transferase Resource Report Resource Website |
Jason Kim | 10.17504/protocols.io.xpmfmk6 | Mouse Metabolic Phenotyping Centers | This experiment involves a spectrophotometric measurement using Roche Cobas Clinical Chemistry Analyzer. Serum alanine transferase levels reflect liver function. | University of Massachusetts | https://mmpc.org/shared/document.aspx?id=180&docType=Protocol | 1 | 2019 | Jason Kim 2019. U Mass - Alanine Transferase. protocols.io dx.doi.org/10.17504/protocols.io.xpmfmk6 | 2021-03-29 03:13:43 | ||
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Vandy - Transverse Aortic Constriction Resource Report Resource Website |
Lin Zhong, Jeffrey Rottman, Chee Lim | 10.17504/protocols.io.6xdhfi6 | Mouse Metabolic Phenotyping Centers | Summary:Hypertension in man can be modeled in the mouse by introducing a constriction in the major artery (aorta) thereby obstructing outflow and increasing the afterload to the heart. This protocol describes the procedure for aortic banding aka transverse aortic constriction (TAC). | Vanderbilt University, Vanderbilt University, Vanderbilt University | https://mmpc.org/shared/document.aspx?id=228&docType=Protocol | 2 | 2019 | Lin Zhong, Jeffrey Rottman, Chee Lim 2019. Vandy - Transverse Aortic Constriction. protocols.io dx.doi.org/10.17504/protocols.io.6xdhfi6 | 2021-03-29 03:13:42 | ||
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Yale - Blood Albumin Resource Report Resource Website |
Gary Cline, John Stack | 10.17504/protocols.io.y2cfyaw | Mouse Metabolic Phenotyping Centers | Procedure used to determine the concentration of albumin in blood, plasma, and serum. Albumin is measured as its conjugate with bromocresol green monitored at 600 nm. | Yale University, Yale University | https://mmpc.org/shared/document.aspx?id=205&docType=Protocol | 1 | 2019 | Gary Cline, John Stack 2019. Yale - Blood Albumin. protocols.io dx.doi.org/10.17504/protocols.io.y2cfyaw | 2021-03-29 03:13:51 |
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