Blood samples were collected and analyzed using the Olympus AU2700 analyzer (Beckman Coulter, High Wycombe, UK) with standard proprietary reagents as follows: glucose with hexokinase, total cholesterol and high-density lipoprotein with cholesterol esterase/oxidase and triacylglycerol with glycerol kinase. Low-density lipoprotein was calculated according to the Friedwald formula. Insulin was measured using radio-immunoassay (Invitrogen, UK). HOMA-IR was calculated using fasting glucose and insulin concentrations.
Au2700 analyzer
Manufactured by Beckman Coulter
Sourced in United States, United Kingdom
The AU2700 analyzer is a fully automated, high-throughput clinical chemistry analyzer designed for routine clinical laboratory testing. It offers a wide menu of assays and can perform multiple types of analyses simultaneously. The AU2700 analyzer is capable of handling a variety of sample types and provides reliable and accurate results.
Automatically generated - may contain errors
Lab products found in correlation
Radioimmunoassay, by Thermo Fisher Scientific (2 mentions)
Mcp 1, by R&D Systems (1 mentions)
Adiva centaur xp analyzer, by Siemens (1 mentions)
Bovine serum albumin (bsa), by MP Biomedicals (1 mentions)
Quantikine mouse epo kit, by R&D Systems (1 mentions)
Mouse il 6 elisa max set deluxe kits, by BioLegend (1 mentions)
Colchicine, by Merck Group (1 mentions)
7 protocols using au2700 analyzer
1
Comprehensive Metabolic Panel Analysis
2
Quantification of Serum and Urine Biomarkers
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The serum and urine concentrations of MCP-1, MCSF, and neopterin were evaluated by commercially available ELISA kits (MCP-1—R&D Systems, reagent kit DCP00; MCSF—R&D Systems, reagent kit DMC00B; neopterin—IBL International, reagent kit RE59321). Standard, serum, and urine samples were transferred to 96-well microplates precoated with recombinant antibodies to human MCP-1, MCSF, and neopterin. Measurements were performed according to the manufacturer's instructions, and results were calculated by reference-to-standard curves.
The serum and urine creatinine were assessed with the Creatinine OSR61204 reagent on the Beckman Coulter AU2700 analyzer.
The fractional parameter excretion was calculated according to the formula ([urine parameter concentration] × [serum creatinine concentration]) / ([serum parameter concentration] × [urine creatinine concentration]) × 100%.
The serum and urine creatinine were assessed with the Creatinine OSR61204 reagent on the Beckman Coulter AU2700 analyzer.
The fractional parameter excretion was calculated according to the formula ([urine parameter concentration] × [serum creatinine concentration]) / ([serum parameter concentration] × [urine creatinine concentration]) × 100%.
3
Glucose Tolerance and Insulin Response
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After an overnight fast, participants underwent a 3-h OGTT with the ingestion of 75 g glucose. Venous blood samples were obtained at 0, 30, 60, 120, and 180 min. Plasma glucose was measured by the hexokinase method using a Beckman AU2700 analyzer (Beckman Coulter, Brea, CA, USA). Serum insulin was assessed by chemiluminescence immunoassay using a Siemens ADIVA Centaur XP analyzer (Siemens Healthcare Diagnostics Inc., Tarrytown, NY, USA). The glucose and insulin assays were standardized to NIST SRM 965 and WHO 1st IRP 66/304, respectively. The repeatability and within laboratory coefficient variations were < 5%.
4
Metabolic Biomarker Analysis Protocol
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Blood samples were collected and analyzed using the Olympus AU2700 analyzer (Beckman Coulter, High Wycombe, United Kingdom) with standard proprietary reagents as follows: glucose with hexokinase, total cholesterol and HDL-cholesterol with cholesterol esterase/oxidase and triacylglycerol with glycerol kinase. LDL-cholesterol was calculated according to the Friedewald formula. Insulin was measured using radioimmunoassay (Invitrogen, Paisley, United Kingdom). HOMA-IR was calculated using fasting glucose and insulin concentrations (Matthews et al., 1985 (link)). The intra- and inter-assay coefficient of variation for all blood parameters was ≤5%.
5
Biomarkers of Renal Function
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Tail blood samples for complete blood count (CBC) and urea were taken two times during experiment and prior to sacrifice. Blood counts were determined using a veterinary heamatology analyzer (Exigo H400, Boule, FL, USA). Tail blood urea was analyzed using an enzyme linked immunosorbent (ELISA) commercial kit (Abcam, Cambridge UK). Serum creatinine, was analyzed using the AU2700 analyzer (Beckman-Coulter, CA, USA), based on the Jaffe method for determination of mouse serum creatinine. Urine albumin was tested by the Coomassie blue reaction on urine samples that were loaded on an SDS-PAGE. Bovine serum albumin (fraction V, MP biomedicals, OH, USA) served as control.
6
Comprehensive Blood and Serum Analyses
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Complete blood count was assessed from tail bleed collected into EDTA-coated capillary tubes (Pro-Vet laboratory, Yodfat, Israel, GENESIS analysis system, Oxford Science, Oxford, USA). Kidney functions and serum iron levels from were analyzed using the AU2700 analyzer (Beckman-Coulter, CA, USA). Serum erythropoietin (EPO) levels were determined by enzyme-linked immunosorbent assays (Quantikine Mouse Epo kit, R&D Systems, Minneapolis, MN. Sensitivity: 47.0 pg/mL; intra- and inter-assay coefficients of variation: < 4.4% and 9.7%, respectively). Serum IL-6 concentrations were performed by mouse IL-6 ELISA Max Set Deluxe Kits (Biolegend, San Diego, CA, USA. Sensitivity: 2 pg/mL).
7
Colchicine impact on CKD mice
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This study was approved by the Ben-Gurion University of the Negev Animal Use and Care Committee, protocol number IL-39-07-2018. All protocols comply with the NIH Guidelines and are reported in accordance with ARRIVE guidelines (https://arriveguidelines.org). Animals were housed in standard laboratory cages. Food and water were given ad libitum. Eight weeks old male C57BL/6 mice (Harlan Laboratories Inc. Rehovot, Israel) were divided into 4 groups (n=6-10 in each group): C, C-col, CKD and CKD-col. CKD was induced by adenine diet [15] . First an 0.3% adenine, 0.9% phosphorus and 75 ppm iron diet was given for 10 days, switched then to a 0.2% adenine diet and unchanged phosphorus and iron, for additional 11 days. Control groups were fed with a control diet (0.3% phosphorus, ~75 ppm iron). All diets were purchased from Envigo Teklad, (Huntingdon, UK). Colchicine (Sigma, Missouri, USA) was diluted in saline to a 10 mg/ml solution and given at a dose of 30 mg/kg intraperitoneally (ip) to C-Col and CKD-Col mice seven days a week, while C and CKD groups were ip injected with saline. Mice were sacri ced after 21 days, using anesthesia with ketamine and xylazine, collecting: blood, liver and kidney. Serum creatinine levels were analyzed using the AU2700 analyzer (Beckman-Coulter, CA, USA).
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