Supernatant from SW‐treated HUVECs was analyzed using a human cytokine array kit as suggested by the manufacturer (human cytokine array kit; R&D Systems, Minneapolis, MN). Pixel density of membranes was quantified using Image J software (NIH, Bethesda, MD).
Human cytokine array kit
Manufactured by R&D Systems
Sourced in United States, Germany, United Kingdom
The Human Cytokine Array Kit is a multiplex assay for the simultaneous detection and quantification of multiple human cytokines, chemokines, and growth factors in a single sample. The kit utilizes capture antibodies spotted on a membrane to bind and detect target analytes. This allows for the parallel measurement of a variety of targets in a simple, rapid, and sensitive manner.
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Lab products found in correlation
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Odyssey imager, by Rockland Immunochemicals (2 mentions)
Irdye 800cw streptavidin, by Rockland Immunochemicals (2 mentions)
Irdye 800cw streptavidin, by LI COR (2 mentions)
Human soluble receptor array kit, by R&D Systems (2 mentions)
Non hematopoietic panel, by R&D Systems (2 mentions)
Chemidoc xrs system, by Bio-Rad (1 mentions)
Chemidoc xrs imaging system, by Bio-Rad (1 mentions)
Cytometric bead array, by BD (1 mentions)
Lsrii cytometer, by BD (1 mentions)
Fcap array v3, by BD (1 mentions)
Legendplex data analysis software, by BioLegend (1 mentions)
Legendplex human m1 m2 macrophage panel, by BioLegend (1 mentions)
Macsquant q10 cytometer, by Miltenyi Biotec (1 mentions)
X ray film, by Fujifilm (1 mentions)
Human il 8 cxcl8 duoset elisa, by R&D Systems (1 mentions)
Human cxcl1 gro alpha duoset elisa, by R&D Systems (1 mentions)
Human phospho kinase array, by R&D Systems (1 mentions)
Human apoptosis array kit, by R&D Systems (1 mentions)
Human angiogenesis array kit, by R&D Systems (1 mentions)
56 protocols using human cytokine array kit
1
Cytokine Profiling of HUVEC Supernatant
2
Cytokine Array Analysis of Chrysophanol Effects
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The cytokine array analysis was performed according to the method of a previous study [30 (link)], with some modifications. Cells were untreated (control) or treated with chrysophanol, and the medium was replaced with serum- and antibiotic-free DMEM for the final 24 h of the incubation. Medium from three wells was pooled, clarified by centrifugation at 200× g at 4 °C, and immediately applied to the Human Cytokine Array Kit according to the manufacturer’s instructions (R&D Systems, Minneapolis, MN, USA). The cytokine array signal was detected at multiple exposure times, ranging from 15 s to 10 min. The film was scanned using a ChemiDocTMXRS + System (Bio-Rad Laboratories, Hercules, CA, USA). Signal levels were measured using ImageJ software, with the Protein Array Analyzer plugin 16.
The Human Cytokine Array Kit was provided by R&D Systems and was used according to manufacturer’s introduction and the method suggested by a previous study [31 (link)]. The image of spots was scanned and measured using NIH ImageJ software with the Protein Array Analyzer plugin 16.
The Human Cytokine Array Kit was provided by R&D Systems and was used according to manufacturer’s introduction and the method suggested by a previous study [31 (link)]. The image of spots was scanned and measured using NIH ImageJ software with the Protein Array Analyzer plugin 16.
3
Cytokine Profiling in Synovial Fluid
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Synovial fluid mediators were determined by screening pro-inflammatory cytokines on three SF using a membrane-based Human Cytokine Array kit (ARY005B, R&D Systems, Minneapolis, MN, USA). The chemiluminescent signals were visualized with a ChemiDoc XRS+ imaging system (Bio-Rad, Hercules, CA, USA) and analyzed using Image Lab 5.0. IL-6 and IFN-γ levels were quantified by ELISA according to the manufacturer's instructions (R&D Systems). Concentrations of MCP-1 levels were determined by Cytometric Bead Array (BD Biosciences, San Diego, CA, USA). Data was acquired on an LSRII cytometer (BD Biosciences) and analyzed using FCAP array v3 software (BD Biosciences). Quantification of IL-1β, IL-23, IL-12p70, IL-12p40, CCL17, CXCL10, IL-10, and IL-1RA in SF was performed by the LegendPlex Human M1/M2 Macrophage Panel (BioLegend, San Diego, CA, USA). Data was acquired on a MACSQUANT Q10 cytometer (Miltenyi Biotec, Bergisch Gladbach, Germany) and analyzed using Legendplex Data Analysis Software (BioLegend).
IL-6 and TIMP3 were quantified in ADSC culture supernatants by ELISA (R&D Systems).
IL-6 and TIMP3 were quantified in ADSC culture supernatants by ELISA (R&D Systems).
4
Cytokine Profiling in Stimulated HT-29 Cells
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To profile the response of stimulated HT-29 epithelial cells, 38 cytokines were simultaneously analyzed using a Human cytokine array kit (Cat no. ARY005, R&D Systems, Abingdon, UK) according to the manufacturer’s instructions. Qualitative analysis was performed by comparing the density of the detected spots to that of the reference spots included in the assay.
5
Cytokine Profiling in Pregnancy
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Cytokines, chemokines, and acute phase proteins were detected with the Human Cytokine Array Kit (R&D Systems) according to the manufacturer’s instructions. Pooled sera collected from control non-pregnant individuals and pregnant donors during the first, second, and third trimesters of gestation were centrifuged and incubated with the pre-coated nitrocellulose membranes. After washing and addition of the detection antibody streptavidin–HRP conjugates, the membranes were exposed to X-ray film (Fuji). The cytokine proteomic array comprised 36 targets spotted in duplicate on the membranes. The intensity of each spot in the captured images was analyzed with ImageJ analysis software (NIH Image Processing).
6
Quantifying Inflammatory Mediators in HaCaT Cells
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Human Cytokine Array Kit (R&D Systems) was used to measure relative levels of inflammatory mediators in cell-free supernatants from HaCaT cells, according to the manufacturer’s protocol. The release of IL-8/CXCL8 and CXCL1/GROα from HaCaT cells was quantified using Human IL-8/CXCL8 DuoSet ELISA and Human CXCL1/GRO alpha DuoSet ELISA (R&D Systems) following the manufacturer’s protocol.
7
Cytokine Array Analysis of Cell Supernatants
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After 24 h incubation, cell supernatants were collected and analyzed using a human cytokine array kit following manufacturer’s instructions (R&D Systems). A pool of at least 12 supernatants per condition of 3 independent experiments were used. The results were normalized to the control condition in each cell type. Data were expressed as a fold change relative to controls.
8
Cytokine Profiling in HK-2 Cells
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CM was collected from HK-2. Relative amounts of cytokine levels were determined using Human Cytokine Array kit (ARY005B, R&D systems) according to the manufacturer’s instructions. CM collected from culture cells were also used for detection of MIF by MIF ELISA kits (BOSTER) according to the manufacturer’s instructions.
9
Profiling Kinase and Apoptosis Proteins in MSCs
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Analysis of phosphorylation profiles of kinases and their protein substrates, as well as analysis of expression of apoptosis-related proteins was done by the Human Phospho-Kinase Array (R&D Systems, Minneapolis, MN) and Human Apoptosis Array Kit (R&D Systems). For both, untreated and overnight 1 μg/ml cisplatin pretreated MSC were solubilized at 1 × 107 cells/ml in lysis buffer at 2–8 °C for 30 min and proceeded according to manufacturer’s protocol. ImageJ software (NIH, Bethesda, MD) was used for the quantitative evaluation; pixel density was determined and calculated.
Cell supernatant of untreated MSC and pretreated MSC as above was analyzed by Human Cytokine Array Kit (R&D Systems) used to simultaneously detect the relative levels of 36 different cytokines, chemokines, and acute phase proteins according manufacturers protocol.
Cell supernatant of untreated MSC and pretreated MSC as above was analyzed by Human Cytokine Array Kit (R&D Systems) used to simultaneously detect the relative levels of 36 different cytokines, chemokines, and acute phase proteins according manufacturers protocol.
10
Cytokine and Adipokine Profiling of BxPC-3 Cells
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