Plasma samples were aliquoted and stored at −20°C until subsequent analysis of the following biomarkers: high‐sensitivity C‐reactive protein (hsCRP), interleukin (IL)‐6, TNF‐α, and soluble CD14 (sCD14). The levels of hsCRP were determined with an immunoturbidimetric serum assay using Cobas 701 (Roche Diagnostics, Mannheim, Germany). Commercially available enzyme‐linked immunosorbent assays were used for the assay of IL‐6 (Quantikine HS Human IL‐6 immunoassay kit; R&D Systems, Minneapolis, MN), TNF‐α (using Quantikine HS Human TNF‐α immunoassay kit; R&D Systems), and sCD14 (Human CD14 ELISA Kit; Thermo Fisher Scientific, Waltham, MA) following the manufacturers' instructions.
Quantikine hs human tnf α immunoassay kit
Manufactured by R&D Systems
Sourced in United States
The Quantikine HS Human TNF‐α Immunoassay Kit is a solid-phase enzyme-linked immunosorbent assay (ELISA) designed to measure human tumor necrosis factor alpha (TNF‐α) concentrations in cell culture supernates, serum, and plasma. The kit utilizes a quantitative sandwich enzyme immunoassay technique.
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3 protocols using quantikine hs human tnf α immunoassay kit
1
Biomarker Assessment in Plasma Samples
2
Body Composition Assessment by DXA
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A DXA scan (Discovery Wi System, Hologic Inc, Bedford, MA, USA) was used to assess the body composition. A 3-compartment model was used for estimating whole body fat, LBM, and mineral density using the software provided by the manufacturer (QDR software version 13.4.1; Hologic Inc, Bedford, MA, USA). Patients were clinically euvolemic at the time of DXA scans. Diuretics were prescribed for patients who presented with edema, and measurements were done when they were euvolemic. The TNF-α level was measured by a commercially available ELISA (Quantikine HS human TNF-α Immunoassay kit, R & D Systems, Minneapolis, MN, USA).
3
Inflammatory Biomarkers Analysis Protocol
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The blood samples for the inflammatory biomarkers were taken under the same procedure, at the same time and place as the samples collected for the analysis of metabolic markers. C-Reactive Protein (CRP) and serum amyloid – A (SAA) were assayed by particle-enhanced immunonephelometry (N Latex, Dade-Behring Marburg GmbH, Marburg, Germany) (49 (link)). Interleukin-6 (IL-6) levels were quantified with high sensitivity enzyme linked immunoassay (R&D System Europe Ltd., Abingdon, UK) and tumor necrosis factor (TNF)-α was measured with ELISA method (Quantikine HS/human TNF-α immunoassay kit, R&D Systems, Inc., Minneapolis, USA) (49 (link)).
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