A sample of 927 individuals from the ALSPAC cohort 24 (link)25 (link), previously genotyped for the HP CNV (see below), was used as the control population. Plasma haptoglobin level was available for 325 of these individuals. It was measured using an immunoturbimetric haptoglobin assay (Cobas Integra kit catalogue number 03005593 322, Roche, USA) on a Hitachi Cobas c311 autoanalyser. In the ALSPAC study, pregnant women resident in Avon, UK with expected dates of delivery 1st April 1991 to 31st December 1992 were invited to take part in the study. Of the 15,247 pregnancies, there were 14,899 children who were alive at 1 year of age. The ALSPAC study website (http://www.bristol.ac.uk/alspac/researchers/our-data/ ) contains details of all the data that is available through a fully searchable data dictionary and variable search tool. Ethical approval for the study was obtained from the ALSPAC Ethics and Law Committee and the Local Research Ethics Committees.
Cobas c311
Manufactured by Hitachi
Sourced in Japan
The Cobas c311 is a clinical chemistry analyzer developed by Hitachi. It is designed for routine diagnostic testing in clinical laboratories. The Cobas c311 has the capability to perform a variety of biochemical tests, including assays for enzymes, metabolites, and other analytes. The device is intended to provide accurate and reliable results to support clinical decision-making.
Automatically generated - may contain errors
Lab products found in correlation
Unicel dxi 800, by Beckman Coulter (2 mentions)
Cobas integra, by Roche (1 mentions)
Emr 500, by Labomed (1 mentions)
Bm 20, by Beurer (1 mentions)
D 78532, by Hettich (1 mentions)
Monolisa hbsag ultra3, by Bio-Rad (1 mentions)
Piercetm bca assay, by Thermo Fisher Scientific (1 mentions)
Cobas c 501, by Hitachi (1 mentions)
10 protocols using cobas c311
1
Haptoglobin Levels in ALSPAC Cohort
2
Vitamin D Replacement and Glycemic Control
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Vitamin D assays were performed using commercially available sandwich ELISA kit, supplied by Chongqing Biospes Co., Ltd (Chongqing, People’s Republic of China) with the catalog number: BYEK1472 using microplate ELISA reader (EMR−500, Labomed, Inc., LA, USA). Vitamin D deficiency was defined as a 25-OHD level of ≤20 ng/ml and vitamin D insufficiency as 21–29 ng/ml and optimal level as ≥30 ng/ml.29 (link)–32 (link)
HbA1c assays were done using cobas c311, Hitachi, Roche Diagnostics, Germany. The patients were considered as poor glycemic control when their HbA1c ˃8%, while those having their HbA1c≤8% were categorized as having good glycemic control.9
Three months later, HbA1C and Vitamin D assays were repeated for vitamin D deficient patients who received vitamin D replacement therapy in addition to the routine insulin therapy.
HbA1c assays were done using cobas c311, Hitachi, Roche Diagnostics, Germany. The patients were considered as poor glycemic control when their HbA1c ˃8%, while those having their HbA1c≤8% were categorized as having good glycemic control.9
Three months later, HbA1C and Vitamin D assays were repeated for vitamin D deficient patients who received vitamin D replacement therapy in addition to the routine insulin therapy.
3
Comprehensive Metabolic and Anthropometric Profiling
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Anthropometric measurements in this study included height (evaluated using a standard height ruler) and weight (measured in an upright position by a calibrated scale). Blood pressure of patients was measured in sitting position by digital devices (BM 20, Beurer, Germany) after 5 minutes of rest. This assessment was repeated two more times, each after 5 minutes of rest. The average of second and third assessments was used for interpretation.
Samples of venous blood were collected from eligible individuals following 12 hours of fasting. Using the autoanalyzer (Cobas C311 Hitachi, Tokyo, Japan), serum was assessed for levels of fasting plasma glucose (FPG), HbA1c, high-density lipoprotein cholesterol (HDL), total cholesterol, and triglycerides (TG). Low-density lipoprotein cholesterol (LDL) was calculated through Friedwald formula if the TG level was less than 400 mg/dL.[17 (link)] All the laboratory samples were transferred to a central laboratory to be assessed by one autoanalyzer to prevent inter-laboratory comparisons bias.
Samples of venous blood were collected from eligible individuals following 12 hours of fasting. Using the autoanalyzer (Cobas C311 Hitachi, Tokyo, Japan), serum was assessed for levels of fasting plasma glucose (FPG), HbA1c, high-density lipoprotein cholesterol (HDL), total cholesterol, and triglycerides (TG). Low-density lipoprotein cholesterol (LDL) was calculated through Friedwald formula if the TG level was less than 400 mg/dL.[17 (link)] All the laboratory samples were transferred to a central laboratory to be assessed by one autoanalyzer to prevent inter-laboratory comparisons bias.
4
Serum Biochemical Analysis in Rats
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Blood samples were collected from all anesthetized rats through cardiac puncture in which the collected blood samples were immediately placed into gel tubes for serum collection, then later centrifuged (Hettich D-78532/Germany) at 3000 rpm for 15 minutes. The sera were stored at −80°C (Sanyo – Ultra – Low Temperature, Moriguchi, Osaka, Japan) for the measurement of creatinine. Serum creatinine, urea and uric acid were determined spectrophotometrically (Cobas c311 Hitachi, Tokyo, Japan) using a kit from BIOLABO S.A.S. (Maizy, France) at 500 nm. Phosphorous was also estimated in serum using the ABNOVA calorimetric kits (Catalaog No. KA0815).
5
Comprehensive Liver and Viral Serology Panel
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Liver biochemistry was analyzed using Hitachi Cobas C311. Hepatitis B serology was performed using an enzyme immunoassay (Monolisa HBsAg Ultra 3; Bio-Rad). Hepatitis C antibody testing was done using 3rd generation enzyme immunoassay (Bio-Rad Monolisa Anti-HCV PLUS). A confirmatory HIV serology test was performed for the HIV-uninfected participants.
6
Inflammatory Biomarkers in Adolescents
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Two inflammatory markers (hsCRP and GlycA) were measured in serum samples at 11–12 years in LSAC (n = 1237) and plasma samples at 15.5 years in ALSPAC (n = 3488). Compared to hsCRP, an acute phase reactant, GlycA is a biomarker of chronic and cumulative inflammation with low intra-individual variability (Connelly et al., 2017 (link)). hsCRP (mg/L) was determined using Roche/Hitachi Cobas c311 and GlycA (mmol/L) was quantified by the high-throughput proton NMR metabolomics (Nightingale Health, Helsinki, Finland) (Clifford et al., 2019 ). hsCRP measurements that were below the detection limit (LSAC: n = 261; ALSPAC: n = 0) were assigned a value equal to 50% of the detection limit (LSAC: 0.01 mg/L). As the distributions of hsCRP and GlycA were skewed, log-transformed hsCRP and GlycA values were used in data analysis, with higher values indicating increased inflammation.
7
Lactate Release Quantification in Cell Culture
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Lactate release in media was performed by colorimetric determination with Cobas® C311 (Hitachi, Tokio, Japan). Therefore growth media from cell culture was centrifuged at 1000 rpm for 10 minutes to remove floating cells and debris and 1.3 ml were removed to a sodium fluoride vessel (Sarstedt, Nümbrecht, Germany). To eliminate influences of phenol red and fetal calf serum, medium was used as negative control and deducted from measurements. Further the lactate content was normalized to intracellular protein concentration to prevent influences of cell numbers and volume. Protein concentration was assessed with PierceTM BCA Assay (Thermofisher Scientific) as described in manufacturer´s instructions.
8
Measuring Fasting Metabolic Markers
9
Glucose and HbA1c Measurement Protocols
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FPG was measured by hexokinase method using the Fasting Glucose Roche/Hitachi Cobas C501 (2011-2014) or the Fasting Glucose Roche Cobas C311 (2015-2018). HbA1c was measured using the A1c G7 HPLC Glycoprotein Analyzer (2011-2012) or the Tosoh Automated Glycoprotein Analyzer HLC-723G8 (2013-2018).
10
Measuring Fasting Metabolic Markers
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Fasting insulin concentrations were measured using one-step immunoenzymatic assay with the Beckman coulter Unicel DXI 800. Fasting plasma glucose (FPG) was measured by a glucose oxidase method. Plasma triglycerides were analyzed on a Roche/Hitachi Cobas c311.
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