Blood samples from the PIVENS trial were collected after a 12 hour fast and, after processing, immediately stored at −70C prior to batch shipping on dry ice to the NIDDK Biosample Repository where they were stored at −80C. Ion mobility analysis (IM) was used to directly quantify the full spectrum of lipoprotein particles in baseline and week 96 serum samples, from the smallest, densest HDL particles to large buoyant VLDL particles, as described previously27 (link). LDL phenotype A was defined as an LDL peak particle diameter ≥ 218.8 Å while LDL phenotype B was defined as an LDL peak particle diameter below ≤ 215.5 Å, with intermediate LDL phenotype I defined as an LDL peak particle diameter between these values. Serum sCD163, ICAM-1, VCAM-1, and E-selectin levels were analyzed in duplicate using the Quantikine© enzyme-linked immunosorbent assay system (R&D Systems, Minneapolis).
Quantikine enzyme linked immunosorbent assay system
Manufactured by R&D Systems
The Quantikine enzyme-linked immunosorbent assay (ELISA) system is a quantitative immunoassay kit designed to measure the concentration of specific proteins or cytokines in biological samples. It utilizes antibody-based detection principles to provide accurate and reproducible results.
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Lab products found in correlation
3 protocols using quantikine enzyme linked immunosorbent assay system
1
Lipoprotein and Biomarker Analysis in NASH
2
Serum Biomarkers in Gastric Bypass
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For all but eight patients, blood draws occurred on the day of gastric bypass surgery when liver tissue was obtained; seven patients had blood drawn within six months before surgery and one patient had blood drawn within one year before surgery. Blood from one 10-ml EDTA-coated tube was separated by centrifugation and serum was stored at −80 °C. Samples were thawed and serum sCD163 and sCD14 levels were batch analyzed in duplicate using the Quantikine enzyme-linked immunosorbent assay system (R&D Systems, Minneapolis, MN). The mean assay coefficient of variance was 3.3% and 4.8% for sCD163 and sCD14, respectively.
3
Quantifying Endotoxin and sCD163 Levels
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Plasma endotoxin (LPS) levels: Samples were diluted with LAL reagent in a ratio of 1:3 and heated at a temperature of 65 °C for half an hour. All samples were analyzed in replica according to the instructions, using the Limulus Amoebocyte Lysate (LAL) assay QCL-1000 (Lonza, Valais Switzerland).
Soluble CD163 (sCD163): Blood samples were collected in a 10-mL EDTA-coated tube, centrifuged and serum was then stored at − 80 °C. The samples were thawed and serum sCD163 was analyzed using the Quantikine enzyme-linked immunosorbent assay system (R&D Systems, Minneapolis, MN). The mean assay coefficient of variance was 3.3% for sCD163.
Soluble CD163 (sCD163): Blood samples were collected in a 10-mL EDTA-coated tube, centrifuged and serum was then stored at − 80 °C. The samples were thawed and serum sCD163 was analyzed using the Quantikine enzyme-linked immunosorbent assay system (R&D Systems, Minneapolis, MN). The mean assay coefficient of variance was 3.3% for sCD163.
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