Blood samples were centrifuged, and sera were stored frozen at −80 °C until assay. We used the Human IL-1 beta/IL-1F2 Quantikine HS ELISA Kit (HSLB00D, R & D Systems, Minneapolis, MN, USA) and Human IL-6 Quantikine ELISA Kit HS (HS600C, R & D Systems, Minneapolis, MN, USA) for measurements of IL-1β and IL-6 in sera. The sensitivity and range of these assay kits were 0.063 pg/mL and 0.1–8 pg/mL, and 0.09 pg/mL and 0.16–10 pg/mL, respectively. IL-1β and IL-6 in serum were measured in duplicate according to the attached protocols. A TriStar LB941-vTi Microplate Reader (Berthold Technologies, Bad Wildbad, Germany) was used to measure optical density (450 nm, reference 560 nm). The standard curves were obtained through cubic regression using the included highly purified E. coli-expressed recombinant human IL-1β and IL-6.
Human il 1 beta il 1f2 quantikine hs elisa kit
Manufactured by R&D Systems
Sourced in United States
The Human IL-1 beta/IL-1F2 Quantikine HS ELISA Kit is a quantitative sandwich enzyme immunoassay designed for the measurement of human interleukin-1 beta (IL-1 beta) levels in cell culture supernates, serum, and plasma. It utilizes a microplate pre-coated with a monoclonal antibody specific for human IL-1 beta.
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3 protocols using human il 1 beta il 1f2 quantikine hs elisa kit
1
Quantification of Serum IL-1β and IL-6
2
Cytokine and Viral Protein ELISA
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Cell culture supernatant was used to perform all ELISAs. Human IL-18 ELISA kit (Abcam, Cambridge, UK), human IL-1 beta/IL-1F2 Quantikine HS ELISA kit (R&D system, Minneapolis, MN, USA) and HIV p24 (ABLinc., Rockville, MD, USA) were utilized following the manufacturer’s recommendations.
3
Influenza Cytokine Response Analysis
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We also measured IL-6 levels in the supernatants because we previously showed that the level of IL-6 production is associated with the extent of influenza virus replication [15] (link). Furthermore, we measured the levels of IL-1β and IL-8 to study the effects of high temperature on the production of inflammatory cytokines following influenza viral infection [15 (link), 39 (link)]. IL-6 levels were measured using a specific enzyme-linked immunosorbent assay (ELISA; a solid phase chemiluminescence ELISA kit, QuantinGlo® ELISA, R&D Systems, MN, USA). IL-1β and IL-8 levels were also measured using a Human IL-1 beta/IL-1F2 Quantikine HS ELISA kit (R&D Systems) and an enzyme immunoassay kit (IL-8 EASIA kit, Invitrogen, CA, USA), respectively.
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