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Human xl cytokine array kit

Manufactured by R&D Systems
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The Human XL Cytokine Array Kit is a quantitative multiplex assay designed to measure the levels of multiple cytokines, chemokines, and growth factors in human cell culture supernatants, serum, plasma, and other biological fluids. The kit utilizes bead-based technology to allow for the simultaneous detection and quantification of up to 102 different analytes from a single sample.

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Close spelling variants of this product

Cytokine arrays (3 citations) Cytokine array kits (2 citations) Human kits (2 citations)

74 protocols using human xl cytokine array kit

1

Cytokine Profiling of ADSC Secretome

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A Human XL Cytokine Array Kit (R&D SYSTEMS., Minneapolis, MN) was used to measure each cytokine secretion of the ADSC’s conditioned medium. The ADSCs were confluented and harvested in each conditioned medium. The procedure used was in accordance with the cytokine assay protocol. Briefly, each membrane was placed in a separate well and incubated with the array buffer for one (1) hour. Samples were prepared by diluting the desired quantity with the array buffer. After aspirating the array buffer and adding prepared samples, they were incubated overnight at 2–8 °C on a rocking platform shaker. Each membrane was washed and added to the diluted detection antibody cocktail then incubated for one (1) hour. After incubation, the prepared Chemi Reagent Mix was inserted onto each membrane. The membrane was exposed to X-ray film and analysed pixel density in each spot of the array.
Wada Y., Ikemoto T., Morine Y., Imura S., Saito Y., Yamada S, & Shimada M. (2019). The Differences in the Characteristics of Insulin-producing Cells Using Human Adipose-tissue Derived Mesenchymal Stem Cells from Subcutaneous and Visceral Tissues. Scientific Reports, 9, 13204.
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2

Cytokine Profiling using XL Array

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Cytokine array was performed by using Human XL Cytokine Array Kit (Cat. no.ARY022, R & D systems, USA) as per the manufacturer’s instructions. In brief, array membrane was first blocked in array buffer 6 for one hour on a rocking platform shaker in a 4 well multi-dish. After blocking, the membrane was incubated with desired sample quantity overnight at 2-8 °C on a rocking platform shaker. Then the membrane was washed for 10 min with 1X wash buffer thrice. 1.5 mL per well of diluted detection antibody cocktail was added and incubated for 1 hour on shaker. Then the membrane was again washed with 1X wash buffer thrice. After washing, membrane was incubated with 2 mL of 1X Streptavidin-HRP for 30 mins. Finally, 1 mL of Chemi Reagent mix was spread evenly on each membrane and autoradiograph was developed. Pixel densities on X-ray films were scanned and analysed using a image analysis software (Image J).
Gangwar A., Paul S., Ahmad Y, & Bhargava K. (2020). Intermittent hypoxia modulates redox homeostasis, lipid metabolism associated inflammatory processes and redox post-translational modifications: Benefits at high altitude. Scientific Reports, 10, 7899.
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3

Profiling Secretome of CAFs and MCF7 Cells

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CAFs and MCF7 cells (as a control) were plated on 10 cm dishes in 10% media. When the cells had reached 80% confluency, media was changed to 1% serum and harvested 48 h later. CAF-CM and MCF7-CM were sterile filtered through 0.22 μm filter and snap frozen in liquid nitrogen or used as fresh. The frozen or fresh CAF-CM and MCF7-CM were then subjected to the Human XL Cytokine Array Kit (R&D ARY022B) according to the manufacturer’s protocol. Quantification of the signal produced by the amount of analyte bound was conducted using Fiji and the data were normalized to MCF7-CM.
Hogstrom J.M., Cruz K.A., Selfors L.M., Ward M.N., Mehta T.S., Kanarek N., Philips J., Dialani V., Wulf G., Collins L.C., Patel J.M, & Muranen T. (2023). Simultaneous isolation of hormone receptor–positive breast cancer organoids and fibroblasts reveals stroma-mediated resistance mechanisms. The Journal of Biological Chemistry, 299(8), 105021.
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4

Cytokine Secretion Profiling of Msh2-deficient Cells

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HCT-116, MFE-296, CT-26 Msh2 KO, and 21B Msh2-null cells were seeded at 5×l05 cells in 6-well tissue culture plates and allowed to adhere overnight. Cells were then treated with either 1 μM MLN4924 or DMSO (control) in 2 mL of growth medium for 24 hours. The supernatants were collected, and the particulates removed by centrifugation. A Human XL Cytokine Array kit (R&D Systems) was used for HCT-116 and MFE-296 cells, whereas a Mouse Cytokine Array Panel A (R&D Systems) measured cytokine secretion from CT-26 Msh2 KO, and 21B Msh2-null cells. Samples were prepared and the ELISAs were processed in accordance with the manufacturer’s instructions. After reference spot normalization, duplicate spots were averaged, followed by subtraction of an averaged background signal. Fold changes between corresponding signals of MLN4924-treated versus DMSO-treated arrays per cell line were calculated. Cytokines represented in both human and mouse arrays were log2-transformed and subjected to unsupervised clustering.
McGrail D.J., Garnett J., Yin J., Dai H., Shih D.J., Lam T.N., Li Y., Sun C., Li Y., Schmandt R., Wu J.Y., Hu L., Liang Y., Peng G., Jonasch E., Menter D., Yates M.S., Kopetz S., Lu K., Broaddus R., Mills G.B., Sahni N, & Lin S.Y. (2020). Proteome instability is a therapeutic vulnerability in mismatch repair deficient cancer. Cancer cell, 37(3), 371-386.e12.
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5

Cytokine Profiling of Cell Lysates

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Cell lysates were collected, and the total protein concentration of each lysate was determined using the BCA method. Lysates from three independent repeats were pooled for the assay. Cytokine assays were conducted with a human XL cytokine array kit (R&D, Minneapolis, MN, USA) according to the standard procedures; the chemiluminescence were detected by ChemiDoc™ Touch Imaging System with Image Lab™ Touch Software 6.1 (Bio-Rad, Hercules, CA, USA). Images were analyzed by HLImage++ software 6.2 (Western Vision Software, Salt Lake City, UT, USA).
Wang L., Su Y, & Choi W.S. (2021). Melatonin Suppresses Oral Squamous Cell Carcinomas Migration and Invasion through Blocking FGF19/FGFR 4 Signaling Pathway. International Journal of Molecular Sciences, 22(18), 9907.
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6

Profiling Cytokine Secretion in Reovirus-Treated Cells

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Media 48 h after treatment with reovirus and GSK2606414 were collected and centrifuged to remove cells or debris. Initial profiling used the proteome profiler human XL cytokine array kit from R&D Systems (MN, USA). Array results were quantified by densitometry using ImageJ (FIJI v.2). Validation of array findings used the human CXCL10/IP-10 DuoSet and human GM-CSF DuoSet ELISA kits from R&D Systems.
McLaughlin M., Pedersen M., Roulstone V., Bergerhoff K.F., Smith H.G., Whittock H., Kyula J.N., Dillon M.T., Pandha H.S., Vile R., Melcher A.A, & Harrington K.J. (2020). The PERK Inhibitor GSK2606414 Enhances Reovirus Infection in Head and Neck Squamous Cell Carcinoma via an ATF4-Dependent Mechanism. Molecular Therapy Oncolytics, 16, 238-249.
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7

Cytokine Profiling of FW-Stimulated HSC3 Cells

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HSC3 cells were plated on a 10 cm cell culture dish (Greiner, Tokyo, Japan) at a density of 1 × 106 /dish on the day before the experiment. Following stimulation with FW for 30 s, the cells were further cultured for 18 h. The culture supernatants were collected and subjected to cytokine array experiments using the Human XL Cytokine Array kit (R&D systems) according to the manufacture's protocol. Images were taken using ChemiDoc XRS (BioRad, Tokyo, Japan).
Kusunoki M., Sata E., Nishio K., Tanaka T., Nishida T., Sugano N., Sato S, & Asano M. (2018). Acid-electrolyzed functional water induces extracellular matrix metalloproteinase inducer, a possible novel alarmin, secretion from oral squamous cell carcinoma cell lines. International Journal of Medical Sciences, 15(12), 1365-1372.
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8

Cytokine Profiling of Bacterial Infections

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Cytokine screening after whole blood infection (with E. coli K12, S. epidermidis RP62A, S. aureus SH1000 and untreated) for 6 h was measured using the Human XL Cytokine Array kit (R&D Systems, Abingdon, UK; n = 1). The membranes were treated as per manufacturer instructions and were imaged on the BioRad ChemiDoc XRS+. Images were processed using ImageJ online software (version 1.50i; https//imagej.net/ij/index.html (accessed between January 2019 and March 2020).
Chick H.M., Rees M.E., Lewis M.L., Williams L.K., Bodger O., Harris L.G., Rushton S, & Wilkinson T.S. (2024). Using the Traditional Ex Vivo Whole Blood Model to Discriminate Bacteria by Their Inducible Host Responses. Biomedicines, 12(4), 724.
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9

Cytokine and Angiogenic Protein Analysis

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The secretion of cytokines in the condition medium (CM) was measured using Human XL cytokine Array Kit (R&D Systems) while the angiogenic-related proteins were measured in the cell lysate using Human Angiogenesis Array Kit (R&D Systems) following the manufacturer’s instructions. Arrays were imaged by Quantity One software with the ChemiDocTM XRS+ system. Protein signals were quantified using ImageJ software.
Gurung S., Williams S., Deane J.A., Werkmeister J.A, & Gargett C.E. (2018). The Transcriptome of Human Endometrial Mesenchymal Stem Cells Under TGFβR Inhibition Reveals Improved Potential for Cell-Based Therapies. Frontiers in Cell and Developmental Biology, 6, 164.
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10

Cytokine Profiling of HMDM Response to Tumor Supernatant

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HMDMs (3 × 105 cells per well of a 12-well plate) were stimulated with LPS (100 ng/mL) for 24 hours after incubation with tumour cell supernatant (TCS) from SKOV3 cells or with TCS and ONA (30 mM) for 24 hours, followed by the determination of IL-10, IL-12 and TNF-α secretion by means of an ELISA kit (eBioscience, San Diego, CA, USA). The cytokine array kit was purchased from R&D Systems (Human XL Cytokine Array Kit #ARY022, Minneapolis, MN, USA).
Tsuboki J., Fujiwara Y., Horlad H., Shiraishi D., Nohara T., Tayama S., Motohara T., Saito Y., Ikeda T., Takaishi K., Tashiro H., Yonemoto Y., Katabuchi H., Takeya M, & Komohara Y. (2016). Onionin A inhibits ovarian cancer progression by suppressing cancer cell proliferation and the protumour function of macrophages. Scientific Reports, 6, 29588.
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