The IL-33 Quantikine ELISA kit was purchased from R&D Systems (Cat#M3300) and manufacturer instructions were followed. 50μL of either brain homogenate supernatant (ex-vivo assay) or CSF was used as sample volume. 4–5 CSF samples from individual animals were pooled to reach 50μL sample volume. IL-33 standards were serially diluted to a minimum of 30pg/mL. Final values of standards and samples were read on a spectrophotometer at 450nm.
Il 33 quantikine elisa kit
Manufactured by R&D Systems
The 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. It utilizes a microplate pre-coated with a monoclonal antibody specific for IL-33.
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3 protocols using il 33 quantikine elisa kit
1
Quantification of Brain IL-33 via ELISA
2
ELISA Quantification of Cytokines
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Enzyme-linked immunosorbent assay (ELISA) for IL-33 and sST2 in serum was performed using the IL-33 Quantikine ELISA Kit (R&D Systems, Inc.) and the sST2 Quantikine ELISA Kit (R&D Systems, Inc.). The supernatant of the cells was collected and centrifuged at 4°C at 1,000 g for 10 min. IL-4 Quantikine ELISA kit (R&D Systems, Inc.) and IL-6 Quantikine ELISA kit (R&D Systems, Inc.) were used to measure the protein secretion of IL-4 and IL-6 in esophageal adenocarcinoma cells following the manufacturer’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).
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