Ezrmi 13k

Manufactured by Merck Group

Sourced in United States, Germany, Switzerland, Canada, Australia

The EZRMI-13K is a laboratory equipment product manufactured by Merck Group. It functions as a compact and versatile rotary microtome, designed for the precise sectioning of biological and medical samples. The EZRMI-13K allows for the preparation of thin, uniform tissue sections for various microscopic and analytical applications.

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135 protocols using ezrmi 13k

Serum Insulin Levels Determination

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Serum (fasting) was analyzed for circulating insulin levels using an insulin enzyme-linked immunosorbent assay (ELISA) kit (EZRMI-13K; Millipore, Australia). All tissue homogenates for ELISAs were prepared using a lysis buffer (10 mM Hepes, 3 mM MgCl₂, 14 mM KCl, 5% glycerol, and 0.2% IPEGAL) containing PhosSTOP Phosphatase Inhibitor Cocktail (Roche) and EDTA-free Complete protease inhibitor cocktail (Roche). Pancreas, liver, and skeletal muscle were analyzed for insulin levels (EZRMI-13K; Millipore). Data were analyzed using an online software program (https://www.assayfit.com/).

Rossetti G., Ermer J.A., Stentenbach M., Siira S.J., Richman T.R., Milenkovic D., Perks K.L., Hughes L.A., Jamieson E., Xiafukaiti G., Ward N.C., Takahashi S., Gray N., Viola H.M., Hool L.C., Rackham O, & Filipovska A. (2021). A common genetic variant of a mitochondrial RNA processing enzyme predisposes to insulin resistance. Science Advances, 7(39), eabi7514.

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Serum Insulin Quantification in Fasted Mice

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Quantification of serum insulin was performed on samples from 4 h fasted mice fasted using a commercial ELISA kit (EZRMI-13K; Millipore Sigma, Burlington, MA, USA), and were measured according to the manufacturer’s instructions and as described in our previous studies [1 (link)].

Chou C.H, & Barton E.R. (2023). Phosphorylation of AMPKα at Ser485/491 Is Dependent on Muscle Contraction and Not Muscle-Specific IGF-I Overexpression. International Journal of Molecular Sciences, 24(15), 11950.

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Metabolic Biomarkers Assessment Protocol

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Fasting serum was assayed for glucose by spectrophotometry (K606-100; BioVision Inc., Milpitas, CA, USA) and for insulin by ELISA (EZRMI-13K; MilliporeSigma, Burlington, MA, USA). Homeostatic model assessment of insulin resistance (HOMA-IR) was calculated using the formula HOMA-IR = glucose × insulin/400. Fasting serum levels of leptin, adiponectin, resistin, and plasminogen activator inhibitor-1 (PAI-1) were determined by multiplex Luminex assays according to the manufacturer’s instructions (MADKMAG-71K and MADPNMAG-70K-01; MilliporeSigma).

Korman B., Marangoni R.G., Lord G., Olefsky J., Tourtellotte W, & Varga J. (2018). Adipocyte-specific Repression of PPAR-gamma by NCoR Contributes to Scleroderma Skin Fibrosis. Arthritis Research & Therapy, 20, 145.

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Hepatic CER Synthesis Protein Analysis

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To assess whether differences in CER content and turnover were driven by changes in protein content, key proteins involved in CER synthesis were measured via Western blot in whole liver homogenate and selected CER synthases in isolated mitochondria. A list of primary and secondary antibodies is presented in the supplemental data. Total protein was assessed with amido black (0.1%; MilliporeSigma) to control for differences in protein loading and transfer as previously described (50 (link)). Liver-TG content was quantified by enzymatic assay using TG (catalog no.: T2449; Sigma) and free glycerol reagent (catalog no.: F6428; Sigma) following Folch lipid extraction (50 (link)). Serum glucose (catalog no.: 997-03001; Wako), TG (same reagents listed for liver-TG), and FFA (catalog nos.: 999-34691; 995-34791; 994-02891; 990-02991, Wako) were quantified using commercially available enzymatic reagents. Serum insulin concentrations were determined using ELISA (EZRMI-13K; MilliporeSigma).

Mucinski J.M., McCaffrey J.M., Rector R.S., Kasumov T, & Parks E.J. (2023). Relationship between hepatic and mitochondrial ceramides: a novel in vivo method to track ceramide synthesis. Journal of Lipid Research, 64(5), 100366.

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Insulin Secretion from Rat Islets

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Islets were isolated by collagenase digestion of the pancreas. For static insulin secretion, pancreatic islets (4 islets per well) were incubated for 1 h with Krebs-bicarbonate buffer (KBR; (in mmol/L) 115 NaCl, 5 KCl, 2.56 CaCl2, 1 MgCl2, 10 NaHCO3, 15 HEPES), supplemented with 5.6 mmol/L glucose and 0.3% BSA and equilibrated with a mixture of 95% O 2 /5% CO 2 to regulate the pH at 7.4. After preincubation time, this medium was discarded and replaced by fresh KBR, and the islets were incubated for 1 h with 2.8 and 11.1 mM glucose. After 1 h of incubation time, the medium was removed and stored at -20 • C. Insulin levels were measured by rat/mouse insulin ELISA kit (EZRMI-13 K; Millipore-Sigma, St. Charles, MO, USA). Total islet protein was assayed using the Bradford dye method (BRADFORD, 1976) with BSA as the standard curve.

Dos Reis Araujo T., Roberta Rodrigues Muniz M., Lourençoni Alves B., Monali Barreto Dos Santos L., Fernandes Bonfim M., Alves da Silva Junior J., Franciesco Vettorazzi J., Cesar Zoppi C, & Magalhães Carneiro E. (2022). Tauroursodeoxycholic acid improves glucose tolerance and reduces adiposity in normal protein and malnourished mice fed a high-fat diet. Food research international (Ottawa, Ont.), 156.

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Glucose Tolerance and Insulin Sensitivity

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At 20 weeks of age, 12 h-fasted mice received an intraperitoneal (ip) injection of glucose (2 g/kg BW). Blood was collected from the tip of the tail vein, and glycemia was measured using an automatic glucometer (Accu-Chek Performa, Roche Diagnostic, USA), before [time (T) 0] and 15, 30, 60, 120 and 180 min after glucose injection. Insulinemia was also measured at 0 and 30 min by rat/mouse insulin ELISA kit (EZRMI-13 K; Millipore-Sigma, St. Charles, MO, USA). To ipITT test, after fasting for two hours, the baseline glycemia was measured by a glucometer and the mice received, intraperitoneally, 0.75 U/Kg body weight of human recombinant insulin (Humulin, Indianapolis, USA). Blood glucose was monitored at 10, 15, 30, 45 and 60 min after insulin administration.

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Comprehensive Metabolic Profiling Protocol

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Plasma fructosamine levels were measured with a colorimetric kit (K450-100, BioVision Inc). Plasma insulin and C-peptide were measured with mouse/rat ELISAs (EZRMI-13K and EZRMCP2-21K, respectively, EMD Millipore). Plasma NEFAs were measured by enzymatic colorimetric assay (NEFA-HR(2), FUJIFILM Wako). Plasma triacylglycerol (TAG) and cholesterol were measured with colorimetric assays (TR22421 and TR13421, respectively, ThermoFisher Scientific). Plasma ALT and AST concentrations were measured with kinetic spectrophotometric assays (A524-150 and A559-150, respectively, Teco Diagnostics).

Kamm D.R., Pyles K.D., Sharpe M.C., Healy L.N., Colca J.R, & McCommis K.S. (2021). Novel insulin sensitizer MSDC-0602K improves insulinemia and fatty liver disease in mice, alone and in combination with liraglutide. The Journal of Biological Chemistry, 296, 100807.

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Liver Lipid Quantification and Serum Insulin

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Triglycerides and free fatty acids (FFAs) in the liver were analysed according to the manufacturer's protocols (Wako Pure Chemical Industries, Ltd., Osaka, Japan). Blood samples were centrifuged at 3,000 × g at 4°C for 15 min to isolate the serum, and insulin levels were detected using an ELISA kit (EZRMI-13K, EMD Millipore, Billerica, MA, USA).

Li W.D., Xia J.R, & Lian Y.S. (2019). Hepatic miR-215 target Rictor and modulation of hepatic insulin signalling in rats. Molecular Medicine Reports, 19(5), 3723-3731.

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Lipid and Glucose Profiling in Plasma

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Lipids from plasma were measured enzymatically on a Hitachi 917 system (Roche Diagnostics GmbH, Mannheim, Germany) using the cholesterol (Cholesterol CHOD-PAP, 11491458-216), and triacylglycerol (Triglycerides GPO-PAP, 11730711) kit from Roche Diagnostics, and the free cholesterol (Free Cholesterol FS, Ref 113609910930), non-esterified fatty acid (NEFA FS, Ref 157819910935) and phospholipid kit (Phospholipids FS, Ref 157419910930) from DiaSys (Diagnostic Systems GmbH, Holzheim, Germany). Plasma bile acid was measured enzymatically on a Roche Modular P chemistry analyzer (Roche Diagnostica), using the BA kit (Total Bile Acid Assy Kit, 05471605001) from Diazyme (Diazyme Laboratories, Gregg, CA, USA). The fatty acid composition was determined by GC/MS as previously described [47 (link)]. Glucose was measured on Hithachi 917 using the Glucose/HK kit (Roche Diagnostics, Ref 11876899-216). Fasting insulin was measured in two parallels of 10 μL plasma from each rat using a rat/mouse insulin 96 well plate assay ELISA kit (EZRMI-13K) from EMD Millipore (Billerica, MA, USA), according to the manufacturer’s instructions.

Ramsvik M.S., Bjørndal B., Bruheim I., Bohov P, & Berge R.K. (2015). A Phospholipid-Protein Complex from Krill with Antioxidative and Immunomodulating Properties Reduced Plasma Triacylglycerol and Hepatic Lipogenesis in Rats. Marine Drugs, 13(7), 4375-4397.

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Insulin Sensitivity Determination Protocol

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Blood glucose was determined using a glucometer (Roche Diagnostics Scandinavia AB, Bromma, Sweden). Serum insulin was determined in duplicate (ELISA kit #EZRMI‐13K; Merck Millipore). Insulin sensitivity was assessed using the homeostatic model assessment of insulin resistance (HOMA‐IR) 13. HOMA‐IR is equal to fasting glucose (millimoles per liter) multiplied by fasting insulin (milliunits per liter) divided by 22.5 14, 15.

Peris‐Sampedro F., Mounib M., Schéle E., Edvardsson C.E., Stoltenborg I., Adan R.A, & Dickson S.L. (2019). Impact of Free‐Choice Diets High in Fat and Different Sugars on Metabolic Outcome and Anxiety‐Like Behavior in Rats. Obesity (Silver Spring, Md.), 27(3), 409-419.

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