Cytokine levels were quantified from media supernatants using ELISA kits for IL-6 (BD Biosciences) and TNF (PeproTech), according to the manufacturers' instructions.
Elisa kits for il 6
Manufactured by BD
Sourced in United States
ELISA kits for IL-6 are laboratory tools used to detect and quantify the presence of the cytokine interleukin-6 (IL-6) in biological samples. These kits provide a standardized and reliable method for measuring IL-6 levels, which can be useful in various research and diagnostic applications.
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4 protocols using elisa kits for il 6
1
Cytokine Quantification by ELISA
2
ELISA Assays for Inflammatory Cytokines
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ELISA kits for IL-6 (#555220), IL-8 (#555244), IL-10 (#555157) and TNF (#555212) were purchased from BD (Franklin Lakes, NJ, USA) and were conducted according to the manufacturers’ instructions. In 96-well plates, 5000 RASF/well were seeded in 200 µL RPMI-1640 with 10% FBS and grown for 72 h. Then, growth medium was replaced by fresh RPMI (2% FBS), and SFs were primed with TNF (10 ng/mL) for 3 days to induce TRPA1 protein. After that, culture medium was replaced with RPMI (2% FBS) and CBG was added for an additional 24 h. After that, supernatants were collected and analyzed. All samples were run in duplicates.
3
Multiplex Cytokine Analysis by ELISA and CBA
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ELISA kits for IL-6 (#550 799), IL-8 (#555 244 BD Bioscience), EMMPRIN (#DY972) and DPPVI/CD26 (#DY1180 R&D Systems, Minneapolis, MN) were used according to the manufacturer's instructions. IL-6 (# 558 276), IL-8 (# 558 277), IL-1β (# 558 279) and TNF-α (# 560 112) were analyzed simultaneously using the Cytometric Bead Array (CBA) Human Master Buffer Kit (#558 264 BD Biosciences, Sweden), using a FACS Canto II flow cytometer operated with the FACSDiva software. Data were analyzed with FCAP Array software, following the manufacturer's tutorial.
4
HA1-2 Vaccine Immune Responses
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Levels of HA1-2-specific IL-6 and IL-23 cytokines were assayed in splenocytes to evaluate cellular immune responses induced by HA1-2-fliC and PEI adjuvanted vaccines. Splenocytes (2 × 106 cells/mL) from immunized mice were incubated for 24 h at 37°C and 5% CO2 in complete RPMI medium with HA1-2 antigen (5 μg/mL). The culture supernatant was collected and concentrations of cytokines were quantitated by ELISA kits for IL-6 (BD Biosciences) and IL-23 (eBioscience), and the IL-6-producing cells were quantified in splenic lymphocyte cultures using the BD™ ELISPOT set (BD Biosciences) according to the manufacturer’s instructions.
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