Blood was transferred into tubes containing 0.3 mol·L−1 EDTA (10 μL·ml−1 blood) and immediately centrifuged at 4°C for 10 min at 23,000 g. The plasma was stored at –80°C until analysis. Analysis of plasma insulin and glucose has been described previously (Ara et al., 2011 (link)). The Quicki index was calculated as previously described (Katz et al., 2000 (link)). Blood HbA1c was analyzed on a DCA Vantage Analyzer (Siemens Healthcare, NY, USA). Plasma IL-6 and plasma TNF-α were measured with high-sensitivity ELISA kits from R&D Systems (Minneapolis, MN, USA). The plasma IL-18 was analyzed with HS ELISA (R&D Systems) and CRP using a conventional high-sensitivity assay on a Cobas Hitachi (Roche, Indianapolis, USA). The plasma adiponectin concentration was determined with a human radioassay kit (EMD Millipore. Billerica, MA, USA). The plasma leptin concentration was measured by a high-sensitivity human ELISA kit (Human leptin Immunoassay, R&D systems).
Hs elisa
Manufactured by R&D Systems
Sourced in United States
The HS ELISA (High Sensitivity Enzyme-Linked Immunosorbent Assay) is a laboratory equipment used for the quantitative detection and measurement of specific proteins or analytes in biological samples. It employs an immunoassay technique that utilizes antibodies and color change to identify and quantify the target molecule of interest.
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2 protocols using hs elisa
1
Blood Biomarkers and Insulin Resistance
2
Inflammatory Biomarkers in Sputum and Serum
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Immunoglobulin (Ig) and albumin concentrations (Abcam) in sputum as well as IL-8, MMP-9 (HS ELISA, R&D Systems), specific IgG against Haemophilus Influenzae type B (HiB) and Streptococcus pneumoniae (PCP: Pneumococcal capsular polysaccharide) (BL International) and IgG subclass levels (Invitrogen) in sputum supernatant as well as serum were quantified using commercial ELISA kits. A Cytokine Multiplex assay was used to simultaneously detect Schnell, 9 the levels of cytokines (IL-1β, IL-5, IL-6, IL-13, TNFα) in sputum supernatant and serum (HS Cytokine premixed panel B, R&D Systems). Assay limits of quantification of the respective analytes are shown in Supplementary Table 1. The formula for albumin correction was calculated as the ratio of sputum analyte value/sputum albumin.
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