After 12 h of LPA stimulation, supernatant from both MDA-MB-231 and MDA-MB-468 cells were collected. Human IL-8 and IL-11 ELISA kit (R&D systems, Minneapolis, MN) were utilized for a quantitative measurement of cytokines according to the manufacturer's recommendations.
Human il 8
Manufactured by R&D Systems
Sourced in United States
Human IL-8 is a cytokine that is primarily produced by macrophages and other cell types. It is involved in the inflammatory response and plays a role in the recruitment and activation of neutrophils.
Automatically generated - may contain errors
Lab products found in correlation
Mouse kc, by R&D Systems (2 mentions)
Mouse il 6, by R&D Systems (2 mentions)
Il 11 elisa kit, by R&D Systems (1 mentions)
Human interferon gamma ifn γ, by Thermo Fisher Scientific (1 mentions)
Dexamethasone dxm, by Merck Group (1 mentions)
Ab11876, by Abcam (1 mentions)
Cytoselect cell migration assay kit, by Cell Biolabs (1 mentions)
P iκb, by Cell Signaling Technology (1 mentions)
Lipopolysaccharide lps from e coli 055 b5, by Merck Group (1 mentions)
P jnk jnk, by Cell Signaling Technology (1 mentions)
Human il 6 elisa kit, by Elabscience (1 mentions)
Inf γ, by R&D Systems (1 mentions)
Neutrophil elastase, by Cell Signaling Technology (1 mentions)
Histone h3, by Cell Signaling Technology (1 mentions)
Gapdh, by Cell Signaling Technology (1 mentions)
Dexamethasone (dex), by Merck Group (1 mentions)
Il 1β, by R&D Systems (1 mentions)
Il 10, by R&D Systems (1 mentions)
Isotype control, by BioLegend (1 mentions)
Sytox red, by Thermo Fisher Scientific (1 mentions)
13 protocols using human il 8
1
Cytokine Quantification in Breast Cancer Cells
2
Gonococcal Neutrophil Infection Assay
3
Cytokine and Inhibitor Regulation of Cellular Processes
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Human interleukin‐1‐beta (IL‐1β), tumour necrosis factor‐alpha (ΤΝF‐α) and human IL‐8 were purchased from R&D Systems, Inc. Human interferon‐gamma (IFN‐γ) was purchased from PeproTech (Rocky Hill, NJ, USA). The NF‐κΒ inhibitor pyrrolidinethiocarbamate ammonium (PDTC) was purchased from TOCRIS Biosciences (Bristol, UK). Dexamethasone (DXM) was purchased from Sigma‐Aldrich (St Louis, MO, USA); bone morphogenetic protein 2 (BMP2), BMP4 and BMP7 were purchased from R&D Systems, Inc.
4
Neutrophil Migration Assay
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Human neutrophils were isolated from peripheral blood of healthy adult donors using Cytoselect Cell Migration Assay kit (Cellbiolabs Inc., San Diego, CA). Neutrophils were suspended in HBSS at the concentration of approximately 1.5×106 cells/ml and 300 µl were loaded into the top chamber of a 24-well Boyden chamber plate (Cell Biolabs inc., San Diego, CA). Sterile PBS or a bacterial suspension (1.5×107 CFU/ml) in chemotaxis medium was added to the bottom chamber. In some experiments, the complement inhibitor Futhan (6-amidino-2-naphthyl p-guanidinobenzoate dimethanesulfonate) [34] (link) was added to the bottom chamber at a final concentration of 50 µg/ml. Human IL-8 (100 ng/ml, R&D Systems) or fMLP 10 nM (N-Formyl-L-methionyl-L-leucyl-L-phenylalanine, 10 nM) (Sigma, St. Louis, MO) were also added to the bottom chamber in some experiments. In some experiments, PBS (vehicle control) or mouse monoclonal anti-human C5a antibody (ab11876, Abcam) dissolved in PBS was added to the bottom chamber at a final concentration of 10 µg/ml. The plates were incubated for two hours at 37°C in tissue culture incubator. The cells that migrated to the bottom chamber were counted using a hemocytometer. Each experiments was repeated with cells from at least 3 different donors.
5
Dehydrozingerone Attenuates Inflammatory Response
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Dehydrozingerone (> 99% purity) bought from TCI chemicals and structure of the compound was shown in Fig. S1A, Lipopolysaccharide (LPS from E. Coli 055: B5) purchased from Sigma Aldrich (USA). Dexamethasone was purchased from Sigma-Aldrich, USA. C-DNA synthesis kit, SYBR green mix purchased from Takara bioscience INDIA. Antibodies GAPDH, JNK P-JNK, c-JUN, p-NF-κB, NF-κB, p38, pP38, Histone-H3, IκB, p-IκB, MPO, Neutrophil Elastase were purchased from Cell signalling technology (USA), ELISA kits (IL-6, CCL2, IL-10, INF-γ, Human-IL-8 and IL-1β were bought from R&D systems. Human IL-6 ELISA kit was procured from Elabsciences (Houston, USA). Secondary Antibodies were procured from Jackson Immuno research private limited (USA). TUNEL kit was procured from Merk Millipore.
6
Detailed Immunological Reagents Inventory
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Sources of Ab for the indicated applications were as follows: for flow cytometry, Ly6G-PE, F4/80-PE, Gr1-PE, CD11b-peridinin-chlorophyll-protein (PerCP), CD115-PerCP, CD244.2-PerCP, CD11b-allophycocyanin, and isotype controls (BioLegend); for immunohistochemistry, CD31, Ly6G, neutrophil elastase, human nucleoli, cit-H3 (Abcam), PNAd (Santa Cruz Biotechnology), MPO (R&D Systems), isotype controls (Thermo Fisher Scientific), and Alexa Fluor 488– and Alexa Fluor 594–conjugated secondary Ab (Life Technologies); and for neutralization, Ly6G (clone 1A8), isotype controls (BioXcell), and human IL-8, MCP-1, GROα, pan-GRO, and G-CSF (R&D Systems). Sources of chemicals were as follows: Sytox Green and Sytox Red (Life Technologies); PMA and DAPI (Sigma-Aldrich); CI-amidine and GSK484 (Cayman Chemical); recombinant DNase I and collagenase type II (Worthington Biochemical Corp.); and D-luciferin (Gold Biotechnology).
7
Cytokine Secretion in Pancreatic Cancer Cells
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MIA PaCa-2, SW-1990 and BxPC-3 cells were seeded at 4×105 cells/well in six-well plates containing medium (MIA PaCa-2 and SW-1990 cells were cultured in DMEM and BxPC-3 cells were cultured in RPMI medium) and cultured for 1 day at 37°C. The culture medium was subsequently changed, and the cells were cultured for an additional 72 h at 37°C in the presence of different concentrations of 10Z-Hymenialdisine (0, 1, 2 and 5 µM). After incubation, the culture medium was collected and centrifuged at 400 × g for 5 min at 14°C to remove particles. The supernatants were frozen at −80°C until use. The concentrations of IL-8 and VEGF were determined using human IL-8 (R&D Systems, Inc.; cat. no. D8000C) and human VEGF (R&D Systems, Inc.; cat. no. DVE00) ELISA kits according to the manufacturer's protocol.
8
SP-A Modulates IL-6 Levels in Murine BAL
9
Multiplex ELISA Quantification
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Mouse MIP-1α/CCL3 (#DY450), mouse KC (#DY453–05), mouse IL-6 (#DY406), human IL-8 (#DY208–05) (R&D Systems, Minneapolis, USA), and mouse albumin (#E99–134, Bethyl Laboratory, Waltham, USA) were quantified by ELISA, and other mediators by 22-plex ProcartaPlex mouse basic kit (Thermo Fisher Scientific, Waltham, USA) on a Luminex-200 instrument (BioRad, Hercules, USA), all according to manufacturer’s instructions.
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Multiplex ELISA Kits for Cytokine Quantification
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