Supernatants collected after miR-338-5p or control miRNA transfections in THP-1 cells were used to perform ELISA for secreted IL-6 using the human IL-6 quantikine ELISA kit (D6050) according to the manufacturer’s protocol (R&D Systems; Minneapolis, MN).
Human il 6 quantikine elisa kit d6050
Manufactured by R&D Systems
Sourced in United States
The Human IL-6 Quantikine ELISA Kit D6050 is a quantitative sandwich enzyme-linked immunosorbent assay (ELISA) designed to measure human interleukin-6 (IL-6) levels in cell culture supernates, serum, and plasma. It utilizes a microplate pre-coated with a monoclonal antibody specific for human IL-6.
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5 protocols using human il 6 quantikine elisa kit d6050
1
Quantification of Secreted IL-6
2
Senescent Cell Secretome Analysis
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Replicative senescent cell models were prepared as described above, and the medium was changed to serum-free medium containing antibiotics. After 24 h, the conditioned medium was collected and IL-6 and IL-8 expression was quantified using ELISA (Human IL-6 Quantikine ELISA Kit (D6050), Human IL-8 Quantikine ELISA Kit (D8000C), (R&D Systems, Inc., Minneapolis, MS, USA)), according to the manufacturer’s protocol.
3
VEGF-A and IL-6 Quantification in A549 Cells
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The expression of VEGF-A and IL-6 in the supernatants of A549 cells was measured with an ELISA for human VEGF and for human IL-6 (Human VEGF Quantikine ELISA Kit DVE00 and Human IL-6 Quantikine ELISA Kit D6050, R&D Systems GmbH, Wiesbaden-Nordenstadt, Germany). The assays are both usable for cell culture supernatants in according to the recommendations of the company. The minimal detectable VEGF concentration is less than 5 pg/ml and the minimal detectable IL-6 concentration is less than 0.70 pg/ml. The VEGF and IL-6 concentrations were then calculated in pg/ml/100,000 viable cells.
4
Oxidative Stress Biomarkers in Acute Ischemic Stroke
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Oxidative stress was analyzed because oxidative stress is an important risk factor for the development of AIS. The serum activity of SOD was measured by the formazan-WST (water-soluble tetrazolium salt) method [52 (link)]. The serum concentration of GSH was determined by using Dithiobis-2-nitrobenzoic acid (DTNB) [53 (link)]. The serum concentrations of AST and ALT were evaluated by using the Hitachi 7170A/7180 Biochemical Analyzer (Hitachi, Japan). Serum levels of IL-6 and TNF alpha were measured by using Human IL-6 Quantikine ELISA Kit D6050 and Human TNF-alpha Quantikine ELISA Kit DTA00C ELISA kits (R&D Systems Inc., Minneapolis, MN, USA), respectively.
5
Quantifying IL-6 Expression in EGFR-Mutant NSCLC
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To check the expression levels of IL-6 protein in 10 EGFR-mutant NSCLC cell lines [16, 17] , the supernatant IL-6 levels in the cell culture were measured by the enzyme-linked immunosorbent assay (ELISA), using Human IL-6 Quantikine ELISA Kit D6050 (R&D Systems, MN, USA). Sensitivities of each NSCLC cell to gefitinib and anti-IL-6 antibody (Biolegend, CA, USA) were evaluated by MTT assay.
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