General laboratory parameters and CSF TP content were measured by spectrophotometry with a Cobas Mira Analyzer (Roche) at the Clinical Biochemistry Lab of the Veterinary Clinical Hospital, UAB. Hemograms were analyzed with a SCIL VetABC hematology analyzer and CSF cell counts, WBC and RBC, were manually counted with a Neubauer chamber by the Clinical Hematology Lab.
Cobas mira analyzer
Manufactured by Roche
Sourced in Switzerland, Germany, United States
The Cobas Mira analyzer is a clinical chemistry system designed for in vitro diagnostic testing. It is capable of performing a variety of automated clinical chemistry analyses, including photometric and ion-selective electrode measurements. The Cobas Mira is intended for use in clinical laboratories and hospital settings to aid in the diagnosis and monitoring of various medical conditions.
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Lab products found in correlation
Abx pentra 400, by Horiba (1 mentions)
Smta00b, by R&D Systems (1 mentions)
Mrp300, by R&D Systems (1 mentions)
Sm6000b, by R&D Systems (1 mentions)
S6050, by R&D Systems (1 mentions)
Elecsys analyzer, by Roche (1 mentions)
Nefa hr 2, by Fujifilm (1 mentions)
S monovette, by Sarstedt (1 mentions)
Glycerol, by Randox (1 mentions)
Rat mouse insulin elisa kit, by Crystal Chem (1 mentions)
Colorimetric assay kit, by Cayman Chemical (1 mentions)
Enzyme immunoassay kit, by Cayman Chemical (1 mentions)
Quantiglo, by R&D Systems (1 mentions)
Ripa buffer, by Merck Group (1 mentions)
Isoproterenol, by Merck Group (1 mentions)
Norepinephrine, by Merck Group (1 mentions)
Fatty acid free bsa, by Merck Group (1 mentions)
Accu check aviva blood glucose monitor, by Roche (1 mentions)
Multi analyte elisarray kit, by Qiagen (1 mentions)
Vacutainer plus plasma tubes, by BD (1 mentions)
20 protocols using cobas mira analyzer
1
Spectrophotometric Analysis of CSF Parameters
2
Standardized Cardiovascular Risk Profiling
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As previously described,8 , 9 BP was measured between 7:00 and 10:00 am using a mercury sphygmomanometer with the auscultatory method, according to recommendations from the seventh report of the Joint National Committee for the Diagnosis, Evaluation, and Treatment of High BP.15 Venous blood was obtained after at least 8 hours of fasting. Total cholesterol, glucose, and triglycerides were measured in serum using enzymatic automated methods (Cobas Mira analyzer; Roche, Basel, Switzerland). Low‐density lipoprotein‐cholesterol and high‐density lipoprotein‐cholesterol were measured by direct methods using the same analyzer. Diabetes mellitus was defined as fasting blood glucose ≥126 mg/dL or pharmacological treatment with glucose‐lowering drugs. Glomerular filtration rate was estimated with the Modification of Diet in Renal Disease equation.16
3
Biochemical Assays for FBS and hs-CRP
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FBS was calculated with a Cobas MIRA analyzer (Roche Diagnostic, Basel, Switzerland) by an enzymatic process (Pars Azmon Co., Tehran, Iran). The sensitivity of the assays for FBS was 5 mg/dL. hs-CRP was calculated by using the turbidimetric method the Pars Azmoon kit (Pars Azmoon Inc., Tehran, Iran) on Hitachi 917. The sensitivity of the assays for hs-CRP was 0.1 mg/L.
4
Bariatric Surgery Blood Lipid Analysis
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In bariatric surgery patients, blood samples were drawn in the morning in the fasting state from the antecubital vein into EDTA and serum tubes (1.6 mg/ml) and centrifuged immediately after collection at 3000 rpm for 10 min at 4 °C. Plasma samples were stored at −80 °C until being used for assaying.
Serum and plasma parameters were measured according to standards as defined by the Central Laboratory Unit of the Medical University of Innsbruck. Plasma triglycerides, total cholesterol, and high-density lipoprotein cholesterol (HDL‑C) were quantified using a commercially available enzymatic kit (Roche Diagnostic Systems, Basel, Switzerland). Low-density lipoprotein cholesterol (LDL‑C) was calculated using the Friedewald formula. Plasma glucose was measured by the hexokinase method on a Cobas MIRA analyzer (Roche Diagnostic Systems, Basel, Switzerland).
Serum and plasma parameters were measured according to standards as defined by the Central Laboratory Unit of the Medical University of Innsbruck. Plasma triglycerides, total cholesterol, and high-density lipoprotein cholesterol (HDL‑C) were quantified using a commercially available enzymatic kit (Roche Diagnostic Systems, Basel, Switzerland). Low-density lipoprotein cholesterol (LDL‑C) was calculated using the Friedewald formula. Plasma glucose was measured by the hexokinase method on a Cobas MIRA analyzer (Roche Diagnostic Systems, Basel, Switzerland).
5
Genotyping of NYD-SP18 SNP in Blood Cells
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Genomic DNA was extracted from the patients' peripheral white blood cells. The NYD-SP18 SNP rs6971091 polymorphism was genotyped using polymerase chain reactionrestriction fragment length analysis. The chemicals used were purchased from Fermentas International Inc., Burlington, Ontario, Canada, and the Polymerase Chain Reactions (PCR) were performed using the DYAD Disciple PCR machine from MJ Research (Waltham, MA, United States). DNA was amplified in a total volume of 25 μL using the oligonucleotides 5´ cct tgg tca tta gct gaa tga gaa gct and 5´ aag gcc tta acc tgg ttc tgc. The PCR product (105 bp) was cut with 5 units of the HindIII restriction enzyme. Restriction fragments of 79 bp and 26 bp represented the A allele, whereas the presence of an uncut product represented the more common G allele.
Plasma triacylglycerols, total cholesterol and cholesterol in high-density (HDL-C) and low-density (LDL-C) fractions were measured enzymatically by standardised procedure, using the Cobas Mira analyzer (Hoffman-LaRoche).
Plasma triacylglycerols, total cholesterol and cholesterol in high-density (HDL-C) and low-density (LDL-C) fractions were measured enzymatically by standardised procedure, using the Cobas Mira analyzer (Hoffman-LaRoche).
6
Lipid and Glucose Biomarker Analysis
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After blood draw and subsequent centrifugation of samples, they were immediately analyzed or stored at −80 °C. Standard laboratory methods using an ABX Pentra 400 (HORIBA ABX SAS, Montpellier, France) were applied for assessment of lipid parameters, triacylglycerol (TG), total cholesterol (TC) and HDL cholesterol (HDL-C). The Friedewald formula (FW) was used for calculation of LDL cholesterol (LDL-C). With a TG level above 3.5 mmol/L, this was not applicable in one subject for which LDL-C is thus missing. A Cobas Mira® Analyzer (Roche Diagnostics, Mannheim, Germany) was used for measurement of glucose in fluoride plasma (S-Monovette® GlucoEXACT; Sarstedt, Nuembrecht, Germany). Serum insulin was analyzed by commercial enzyme-linked immunosorbent assay (Mercodia, Uppsala, Sweden; intra-assay CV 2.8–4.0%, inter-assay CV 4–5%).
7
Metabolic Assessments in Fasting Mice
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Mice were fasted overnight before metabolic analysis, and blood samples were drawn from the retro-orbital plexus of the fasting mice under anesthesia. We used ELISAs to determine serum levels of insulin (ALPCO; 80-INSMSU-E01), leptin (ALPCO; 22-LEPMS-E01), and adiponectin (R&D Systems; MRP300). We used a colorimetric/fluorometric kit to determine serum and tissue levels of NEFAs (BioVision; K612) and ELISA kits to determine serum and tissue levels of TNF-α (R&D Systems; SMTA00B) and IL-6 (R&D Systems; SM6000B and S6050). We measured serum cholesterol and TG levels with a Cobas Mira analyzer (Roche Diagnostics) at the Mouse Metabolic Phenotyping Center in Yale University.
For glucose tolerance tests (GTTs), we determined basal fasting glucose levels and then administered dextrose (1 g/kg body weight) by i.p. injection. For ITTs, we administered insulin (0.35 U/kg body weight for NCD-fed mice, 0.6 U/kg body weight for HFD-fed mice) by i.p. injection. We obtained blood samples from the tail vein 15, 30, 60, and 120 min after injection and determined blood glucose levels using a glucometer (LifeScan).
For glucose tolerance tests (GTTs), we determined basal fasting glucose levels and then administered dextrose (1 g/kg body weight) by i.p. injection. For ITTs, we administered insulin (0.35 U/kg body weight for NCD-fed mice, 0.6 U/kg body weight for HFD-fed mice) by i.p. injection. We obtained blood samples from the tail vein 15, 30, 60, and 120 min after injection and determined blood glucose levels using a glucometer (LifeScan).
8
Lipid and Inflammatory Biomarker Assays
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Plasma insulin was measured using a specific electrochemiluminescence immunoassay for the Elecsys analyzer (Roche Diagnostics, Mannheim, Germany), with a coefficient of variation (CV) of 1.1%. Total cholesterol (CV: 1.0%), triglyceride (CV: 0.9%), glucose (CV: 0.8%), and plasma uric acid (CV: 0.9%) concentrations were measured enzymatically with kits and calibrators supplied by Roche Diagnostics on a Cobas Mira analyzer (Roche Diagnostics). High-density lipoprotein (HDL; CV: 1.5%) was measured in the supernatant after precipitation of apolipoprotein B-containing lipoproteins with phosphotungstate/magnesium chloride solution (23 (link)). Low-density lipoprotein (LDL) was calculated using the Friedewald equation (24 (link)). High sensitivity C-reactive protein (CRP) was measured using a latex-enhanced immunoturbidimetric method (Roche Diagnostics) with a CV of 6.4%.
9
Equine Plasma NEFA Quantification
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Mare plasma NEFA concentrations were measured in duplicate with an enzymatic-colorimetric method using a Cobas Mira-analyzer (Roche, Mannheim, Germany) with a commercial kit for NEFA (NEFA-HR(2), Wako Chemical GmbH, Neuss, Germany). The minimum level of detection was 10 µmol/L. Intra- and inter-assay coefficients of variation were 2.7% and 4.5%, respectively.
10
Plasma Lipid Profile Measurement
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Plasma cholesterol, HDL‐C and triglyceride levels were measured using a COBAS Mira analyzer (Roche Diagnostics, Indianapolis, IN, USA). LDL‐C was calculated using the Friedewald formula. Pre‐β HDL levels were measured using the commercially available ELISA kit by Daiichi Pure Chemicals (Tokyo, Japan).
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