Plasma IL-33 level was measured using a technique described previously [12 (link)]. Briefly, the blood samples were centrifuged (10 min at 1,000×g) within 30 min after collection in tubes containing sodium ethylenediaminetetraacetic acid anticoagulant, and the collected plasma was stored at ≤ − 20 °C. Plasma samples were prepared for analysis in a 96-well plate utilizing a custom human cytokine. IL-33 level was measured by enzyme-linked immunosorbent assay (ELISA) using a Human IL-33 Quantikine ELISA Kit (R&D Systems, Minneapolis, MN, USA) and 12 times concentrated with Amicon Ultra-0.5 mL Centrifugal Filter (Merck Millipore, Burlington, MA, USA) devices to improve the detection yield. We followed the kit-specific protocol provided with the BioTek’s PowerWave XS analyser (BioTek Instruments, Winooski, VT, USA). The result is described as 450 optical density (OD).
Human il 33 quantikine elisa kit
Manufactured by R&D Systems
Sourced in United States, United Kingdom
The Human IL-33 Quantikine ELISA Kit is a quantitative sandwich enzyme immunoassay designed to measure human Interleukin 33 (IL-33) levels in cell culture supernates, serum, and plasma.
Automatically generated - may contain errors
Lab products found in correlation
Amicon ultra 0.5 ml centrifugal filter, by Merck Group (1 mentions)
Human il 4, by R&D Systems (1 mentions)
Il 33 quantikine elisa kit, by R&D Systems (1 mentions)
Bca assay, by Thermo Fisher Scientific (1 mentions)
Mouse il 18 elisa kit, by MBL Life Science (1 mentions)
Human st2 il 33r duoset elisa, by R&D Systems (1 mentions)
4 protocols using human il 33 quantikine elisa kit
1
Plasma IL-33 Quantification Protocol
2
Quantifying Serum Cytokine Levels
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Serum level of IL-33 was determined with Human IL-33 Quantikine ELISA Kit (R&D Systems). IL-4, IL-6 and IL-13 levels in culture supernatants were also detected with Human IL-4, IL-6 and IL-13 Quantikine ELISA Kits (R&D Systems) respectively according to the manual’s instructions.
3
IL-33 Quantification in Ocular Samples
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IL-33 concentrations in the vitreous, retina lysate, serum, and rMC-1 culture supernatant were measured using the mouse/rat IL-33 Quantikine ELISA kit (R&D Systems). CCL2, IL-1α, IL-1β, and ST2 were quantified with Quantikine ELISA kits. IL-18 was measured with mouse IL-18 ELISA kit (MBL International). Cytokine concentrations in the retina lysate were normalized to total protein content measured by BCA assay (Thermo Fisher Scientific). To assess IL-33 levels in the vitreous of AMD patients, patients diagnosed with AMD (one male and five females, age 68–91, median age 79), macular pucker (three males and nine females, age 56–79, median age 72), and macular hole (5 males and 16 females, age 46–75, median age 65) were followed and operated by a single vitreoretinal surgeon (Midwest Eye Institute) with approval from Western Institutional Review Board (WIRB) and written patient informed consent. Transconjunctival pars plana vitrectomy was performed under local anesthesia using a 25-gauge cannula (Alcon). IL-33 levels in the vitreous were measured using the human IL-33 Quantikine ELISA kit (R&D Systems).
4
Plasma Biomarkers of Inflammation
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Plasma concentrations of IL-33, ST2IL1RL1, and IL1RAcP were measured by enzyme-linked immunosorbent assay (ELISA) using Human ST2/IL-33R DuoSet ELISA, Human IL-1 RAcP/IL-1 R3 DuoSet ELISA and Human IL-33 Quantikine ELISA Kit (R&D systems, Abingdon, U.K), according to the manufacturer’s protocol. The assay ranges for IL-33, ST2/IL1RL1, and IL1RAcP were 3.1–200 pg/mL, 31.2–2000 pg/mL, and 31.2–2000 pg/mL, respectively.
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