The concentrations of plasma IL-37, OPG, and hs-CRP were measured by an enzyme-linked immunosorbent assay (ELISA) according to instructions provided by the manufacturers (IL-37: Adipogen AG, Liestal, Switzerland; OPG: MBL, Nagoya, Japan; and hs-CRP: Siemens Healthcare, Tarrytown, New York). All samples were measured in duplicate.
Il 37
Manufactured by Adipogen
Sourced in Switzerland
IL-37 is a recombinant human protein that functions as an interleukin. It is used for research purposes in life sciences laboratories.
Automatically generated - may contain errors
6 protocols using il 37
1
Quantification of Inflammatory Markers
2
Quantification of Plasma IL-37 Levels
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The level of plasma IL-37 (Adipogen AG, Liestal, Switzerland) was measured by an enzyme-linked immunosorbent assay (ELISA), following the manufacturer's instructions. The minimal detectable concentration of IL-37 by this assay is 10 pg/ml for IL-37. The ELISA intra-assay and inter-assay coefficients of variation are <5% and <10%, respectively. All of the samples were measured in duplicate.
3
Plasma Cytokine Quantification by ELISA
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The levels of plasma IL-37 (Adipogen AG, Liestal, Switzerland), IL-18 (MBL, Nagoya, Japan), and IL-18BP (RayBiotech, Norcross GA, USA) were measured by an enzyme-linked immunosorbent assay (ELISA), following the manufacturer's instructions. The minimal detectable concentrations were 10 pg/mL for IL-37, 12.5 pg/mL for IL-18, and 20 pg/mL for IL-18BP. The ELISA intra-assay and interassay coefficients of variation were <5% and <10%, respectively. All of the samples were measured in duplicate.
4
Quantification of IL-18, IL-37, and IL-18BP
5
Antibody Sourcing for Protein Analysis
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β-Actin antibody was purchased from Santa Cruz Biotechnology (Dallas, Texas, USA), and HRP-conjugated rabbit anti-human IgG was purchased from Jackson ImmunoResearch (West Grove, PA, USA); ICAM-1 and IL-18 antibody were purchased from Abcam (Cambridge, MA, USA). IL-37 was purchased from Adipogen AG, Liestal, Switzerland, IL-18 from MBL, Nagoya, Japan, IL-18BP from RayBiotech, Norcross GA, USA, and hs-CRP from Elisa Biotech (Shanghai, China).
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β-Actin antibody was purchased from Santa Cruz Biotechnology (Dallas, Texas, USA), and HRP-conjugated rabbit anti-human IgG was purchased from Jackson ImmunoResearch (West Grove, PA, USA); ICAM-1 and IL-18 antibody were purchased from Abcam (Cambridge, MA, USA). IL-37 was purchased from Adipogen AG, Liestal, Switzerland, IL-18 from MBL, Nagoya, Japan, IL-18BP from RayBiotech, Norcross GA, USA, and hs-CRP from Elisa Biotech (Shanghai, China).
6
Serum Cytokine Levels in Inflammatory Conditions
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Venous blood was collected in conjunction with routine laboratory testing for each visit, and serum obtained and stored at − 80 °C until analysis, as described46 (link)47 (link). Control serum was obtained from 28 healthy individuals whose age and gender was not significantly different to the patients. All controls had normal ESR, indicating that they represent a ‘non-inflamed’ population.
Specific enzyme-linked immunosorbent assay (ELISA) kits were used to measure serum IL-10 (Biolegend, San Diego, CA, USA) and IL-37 (Adipogen, Liestal, Switzerland) according the manufacturer’s protocols. Each sample was tested in duplicate. Where multiple samples over time were available, a TAM IL-10 or IL-37 concentration was calculated46 (link). Any samples that exceeded the upper limit of the standard curve was subsequently diluted in PBS with 1% BSA and re-tested.
Specific enzyme-linked immunosorbent assay (ELISA) kits were used to measure serum IL-10 (Biolegend, San Diego, CA, USA) and IL-37 (Adipogen, Liestal, Switzerland) according the manufacturer’s protocols. Each sample was tested in duplicate. Where multiple samples over time were available, a TAM IL-10 or IL-37 concentration was calculated46 (link). Any samples that exceeded the upper limit of the standard curve was subsequently diluted in PBS with 1% BSA and re-tested.
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