For all in vivo experiments mouse blood samples were collected via cardiac puncture and isolated plasma was analyzed by the COBAS c311 system (Roche) for plasma lipids.
Cobas c311 system
Manufactured by Roche
Sourced in Germany
The COBAS c311 system is a compact and fully automated clinical chemistry analyzer designed for high-throughput laboratory testing. It is capable of performing a wide range of routine and specialized clinical chemistry assays, including tests for metabolic disorders, liver and kidney function, and therapeutic drug monitoring. The system features an intuitive user interface, efficient workflow, and robust performance to support reliable and efficient laboratory operations.
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Lab products found in correlation
Glycerol kit, by R-Biopharm (1 mentions)
Human insulin specific ria kit, by Merck Group (1 mentions)
Cobas fara, by Roche (1 mentions)
Immulite 1000 system, by Siemens (1 mentions)
Unicel dxi 800 analyser, by Beckman Coulter (1 mentions)
Cobas e411 system, by Roche (1 mentions)
Cobas integra 400 plus, by Roche (1 mentions)
Immulite 1000, by Siemens (1 mentions)
Total bile acid assay kit, by Cell Biolabs (1 mentions)
Infinity cholesterol kit, by Thermo Fisher Scientific (1 mentions)
8 protocols using cobas c311 system
1
Plasma Lipid Analysis in Mouse Model
2
Liver Enzyme Plasma Analysis in Mice
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For all in vivo experiments mouse blood samples were collected via cardiac puncture and isolated plasma was analyzed by the COBAS c311 system (Roche) assay kit for liver enzymes. Data was subjected to a student’s t-test.
3
CRP Quantitation by Immunoassay
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Quantitation of CRP was performed on COBAS C311 system (Roche, Basel, Switzerland) based on immunoturbidimetric assay. CRP level was interpreted as normal if less than 6 mg/L.
4
Individualized Cooling Protocol: Metabolic Markers
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During the individualized cooling protocol, blood was collected at thermoneutrality and during mild cold exposure. Plasma concentrations of glucose (ABX Glucose HK CP, Radiometer, Horiba ABX, Montpellier, France), free glycerol (Glycerol kit; R-Biopharm, Darmstadt, Germany), and total glycerol (ABX Triglycerides CP, Radiometer, Horiba ABX) were determined on a COBAS FARA centrifugal spectrophotometer (Roche Diagnostics, Woerden, the Netherlands). Triglyceride levels were calculated using the difference in total and free glycerol. Plasma catecholamines were determined using reagents from Recipe (Recipe Chemicals and Instruments, München, Germany) and analyzed on a HPLC and by electrochemical detection. Serum insulin was analyzed with a Human Insulin-Specific RIA Kit (Millipore) on a Gamma Counter (2470 Automatic Gamma Counter Wizard; Wallac, PerkinElmer, Waltham, MA). The plasma inflammatory marker C-reactive protein (CRP) was measured with a particle-enhanced immunoturbidimetric assay on a COBAS c311 system (Roche Diagnostics GmbH, Mannheim, Germany), and IL-6 and IL-8 were measured with a chemiluminescent immunometric assay on an IMMULITE 1000 system (Siemens, München, Germany).
5
Comprehensive Metabolic and Hormonal Assessments
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After completing the questionnaire, venous blood samples were collected from the participants between 7:00–10:00 am after overnight fasting. The samples were stored at each centre temporarily and were all tested in the reproductive centre of Tongji Medical College. Serum total testosterone (TT) and sex hormone-binding globulin (SHBG) were tested using a chemiluminescent immunoassay method on a UniCel DxI 800 analyser (Beckman Coulter, Brea, USA). Free testosterone (FT) concentration was calculated by the Vermeulen formula [16 (link)]. The serum concentrations of fasting plasma glucose (FBG), insulin, HDL-C, total cholesterol (TC) and triglyceride (TG) were measured directly with a cobas c 311 system (Roche Diagnostic system) (only samples collected from Hubei were tested for the insulin concentration). A homeostasis model assessment (HOMA) index was calculated according to the formula (FBG [mmol/L] × fasting insulin [mU/L])/22.5 [17 (link)]. The HOMA index was used to evaluate the levels of insulin resistance HOMA-IR.
6
Comprehensive Diagnostic Evaluation Protocol
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All patients underwent a detailed history-assessment, thorough clinical and radiological evaluation, and the following investigations:
Routine laboratory investigations:
The automated haematology analyser Sysmex KX 21N (Kobe, Japan) was used to perform a complete blood count (CBC), coagulation profile using Stago STA (Diagnostica Stago, Paris, France), D-dimer using Immulite 1000 (Siemens Healthcare Diagnostics, New York, USA), and biochemical analyses using the Cobas c311 system (Roche Diagnostics, Mannheim, Germany) for kidney and liver function parameters; for ferritin, use the Cobas e411 system; and for C-reactive protein (CRP), use the Cobas Integra 400 plus.
Human citrullinated histone H3 was measured by ELISA using an ELISA kit supplied by the Bioassay Technology Laboratory (lot No 202203013, Zhejiang, China) in accordance with the manufacturer ‘s directions. The assay had a sensitivity of 1.352 ng/mL with a range of 32–105 ng/mL.
Molecular analyses of TLR2 rs5743708 and TLR9 rs5743836 variants:
7
Bile Acid and Lipid Profiling
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BA levels in plasma were analyzed by LC-MS, and total BAs in feces were measured by using the Total Bile Acid Assay Kit (Cell Biolabs, San Diego, CA, USA) according to manufacturer instructions. Plasma alanine transaminase, aspartate transaminase, HDL cholesterol, LDL-C, and total cholesterol were measured by using Cobas c311 system (Roche Diagnostics). Liver and fecal content of cholesterol and triglycerides was measured from lipid extracts as previously described by using an Infinity Cholesterol Kit (Thermo Fisher Scientific) and LabAssay Triglyceride Kit, respectively.
8
Plasma Lipids and Glucose Tolerance
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Plasma total cholesterol (TC), triglycerides (TG) and phospholipids (PL) were measured with enzymatic methods. Quantitative determination of urea nitrogen in plasma (BUN) and creatinine was assayed by immunoenzymatic method with the Roche Cobas c311 system.
For the glucose tolerance test, mice were injected, after 5 h fast, with 2 g/Kg glucose via i.p. injection of 20% w/v sterile glucose solution in PBS. Glucose levels were determined using the One touch Ultra glucometer every 30 min.
For the glucose tolerance test, mice were injected, after 5 h fast, with 2 g/Kg glucose via i.p. injection of 20% w/v sterile glucose solution in PBS. Glucose levels were determined using the One touch Ultra glucometer every 30 min.
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