The concentrations of IL-18, IL-18BP, and IL-37 in sera and in the QFT culture supernatants were determined using commercial specific enzyme-linked immunosorbent assays (Human Total IL-18 DuoSet (R&D, Minneapolis, USA), Human IL-18BPa DuoSet (R&D) and Human IL-37/IL-1F7 Duoset (R&D), Human IFN-γDuoset (R&D) and Human IP-10 Duoset ELISA (R&D)), and processed according to the manufacturer’s specifications. After measuring the concentrations of both IL-18 and IL-18BP in each sample, the law of mass action was used to calculate the level of free IL-18. Single IL-18BP molecule binds a single molecule of IL-18, that interaction has a dissociation constant (Kd) of 0.4 nM. Therefore, the level of free IL-18 was calculated from the equation where is [IL-18free], b is = [IL-18BP] – [IL-18] + Kd, and c is -Kd × [IL-18] (Migliorini et al., 2010 (link)). The absorbance was measured using the Victor 2 Multi-Label Counter Microplate Reader (Wallac Oy, Turku, Finland).
Human il 18bpa duoset
Manufactured by R&D Systems
Sourced in United States
The Human IL-18BPa DuoSet is a set of matched antibodies designed for the quantitative measurement of human interleukin-18 binding protein alpha (IL-18BPa) levels in cell culture supernatants, serum, and plasma samples. The set includes a capture antibody and a detection antibody, allowing for the development of a sandwich ELISA.
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Lab products found in correlation
Human ifn γ duoset, by R&D Systems (1 mentions)
Human ip 10 duoset elisa, by R&D Systems (1 mentions)
Purelink rna mini kit, by Thermo Fisher Scientific (1 mentions)
Superscript 2 reverse transcriptase, by Thermo Fisher Scientific (1 mentions)
Taqman gene expression assay, by Thermo Fisher Scientific (1 mentions)
Human il 18 elisa kit, by MBL Life Science (1 mentions)
Rneasy micro kit, by Qiagen (1 mentions)
2 protocols using human il 18bpa duoset
1
Quantification of IL-18, IL-18BP, and IL-37 in Sera and Cell Supernatants
2
RNA Isolation, cDNA Synthesis, and Cytokine Quantification
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RNA was purified using the RNeasy Micro kit (Qiagen, Hilden, Germany) for patient material or the PureLink RNA mini kit (Invitrogen, Carlsbad, CA, USA). cDNA synthesis was performed using Super Script II Reverse Transcriptase (Invitrogen), according to manufacturer's protocols. mRNA levels were analyzed in duplicate by qPCR using TaqMan Gene Expression Assays [Applied Biosystems, Foster City, CA, USA; Assay IDs: Hs00171042_m1 (IP-10/CXCL10), Hs00158032_m1 (IDO), Hs00271720_m1 (IL-18BP), Hs01555410_m1 (IL-1β), Hs00174103_m1 (IL-8/CXCL8)] and normalized to housekeeping gene GAPDH. Cytokine levels in plasma were measured by sandwich ELISA according to the manufacturer's protocols (IFN-γ, IL-6, IL-1β High Sensitivity ELISA, eBioscience, San Diego, CA, USA; Human IL-18 ELISA kit MBL International, Woburn, MA, USA; Human IL-18BPa Duoset, R&D Systems, Minneapolis, MN, USA). IP-10, MIG and IL-8 levels were quantified by sandwich ELISA as described [37, 38] . Levels of free IL-18 were calculated as described [39] .
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