Min Tu, Xian Liu, Bei Han, Qianqian Ge, Zhanjun Li, Zipeng Lu, Jishu Wei, Guoxin Song, Baobao Cai, Nan Lv, Kuirong Jiang, Shui Wang Yi Miao Wentao Gao. Vasohibin‑2 promotes proliferation in human breast cancer cells via upregulation of fibroblast growth factor‑2 and growth/differentiation factor‑15 expression. DOI: 10.3892/mmr.2014.2317.

...Quantitative sandwich-based antibody array (#QAH-GF-1; RayBiotech, Guangzhou, China) was used to detect 40 human growth factors in lysates of the MCF7-EGFP, MCF7-VASH2, BT474-scramble and BT474-shVASH2 cells. All detection services were provided by RayBiotech (Norcross, GA, USA). Each antibody produced four dots, and the averages of the median signal intensities were used for all calculations. Fold change of >1.5 or <0.66 compared with controls was selected as the distinction between overexpression or knockdown, respectively...

Nina M Poole, Gayatri Mamidanna, Richard A Smith, Lewis B Coons and Judith A Cole. Prostaglandin E2 in tick saliva regulates macrophage cell migration and cytokine profile. Parasites & Vectors 2013, 6:261 doi:10.1186/1756-3305-6-261.

...RayBio® Mouse Cytokine Antibody Array (Catalog # AAM-CYT-1-8) RayBiotech, Inc., Norcross, GA), which simultaneousldetects 22 cytokines. Macrophages were cultured at a density of 5 × 10 (4) cells/ml in 6-well plates and grown to confluence. Twenty-four hours prior to the experiment, cells were changed to medium containing 2% FBS. Cells were treated with vehicle (DMSO), saliva (2.4 μg protein/ml), 10 μM AH 6809, saliva + AH 6809, 0.76 μg/ml lipopolysaccharide (LPS) (InvivoGen, San Diego, CA), or saliva + LPS for 18 h [61]. LPS is a Gram-negative bacteria toxin which activates macrophages through Toll-like receptor 4 (TLR4). The conditioned medium was collected, and the cytokine content for each sample was determined according to manufacturer’s instructions. Arrays were developed with kit detection buffer and exposed to Classic Blue Autoradiography Film X (Molecular Technologies, St. Louis, MO) for 0.5, 2.5, 1, and 5 min. The intensities of signals for each cytokine were quantified by densitometry using ImageJ version 1.46 Windows (National Institutes of Health, Bethesda, MD...

Wafaa Gaber, Ghada S. Azkalany, Tamer A. Gheita, Abeer Mohey, Randa Sabry. Clinical significance of serum interleukin-6 and −174 G/C promoter polymorphism in Rheumatoid arthritis patients. The Egyptian Rheumatologist, Volume 35, Issue 2, April 2013, Pages 107–113 http://dx.doi.org/10.1016/j.ejr.2012.11.002

...Serum IL-6 was assayed using Human IL-6 ELISA kit (quantitative sandwich enzyme immunoassay technique), provided by Ray Biotech, Inc...

Provençal N, Suderman MJ, Vitaro F, Szyf M, Tremblay RE (2013) Childhood Chronic Physical Aggression Associates with Adult Cytokine Levels in Plasma. PLoS ONE 8(7): e69481. doi:10.1371/journal.pone.0069481

...Blood samples (8 to 10 ml) were drawn in EDTA coated-tubes and shipped on ice to the laboratory within 24 hours. Blood samples were immediately resuspended in equal volume of PBS (1X) and plasma isolation was performed by centrifugation on a Ficoll-Paque gradient (GE healthcare). The samples were then frozen and stored at −80°C in aliquots of 1 ml. Quantification of 10 cytokines in plasma was done by ELISA using the Quantibody® Human Inflammation Array 1 from RayBiotech following the manufacturer instructions for both time points...

Korkaya et al., Activation of an IL6 Inflammatory Loop Mediates Trastuzumab Resistance in HER2+ Breast Cancer by Expanding the Cancer Stem Cell Population, Molecular Cell (2012), http://dx.doi.org/10.1016/j.molcel.2012.06.014.

...The equal numbers of cells were plated and cultured for 3 days. Subsequently, conditioned media from these cell cultures were collected and analyzed by the RayBio Human Cytokine Antibody Array 5 (RayBiotech, Inc. Norcross, GA). ELISA assay was performed using the conditioned medium collected from two day cultures of cells seeded at 200,000 cells/plate. Blood samples were drawn through orbital vein just before sacrificing the mice. Plasma separated from whole blood by centrifugation at 14,000 rpm at 40C. Plasma from tumorbearing mice and the conditioned medium from in vitro cultures were then analyzed for the indicated cytokines by UM Cytokine Core facility...

Damayanti Chakraborty, M. A. Karim Rumi, Toshihiro Konno, and Michael J. Soares. Natural killer cells direct hemochorial placentation by regulating hypoxia-inducible factor dependent trophoblast lineage decisions. Proc Natl Acad Sci U S A. 2011 Sep 27; 108(39): 16295–16300. Published online 2011 Sep 7. doi: 10.1073/pnas.1109478108 PMCID: PMC3182743 Developmental Biology.

...VEGFA was measured by ELISA (RayBiotech)....

Rundqvist H, Augsten M, Strömberg A, Rullman E, Mijwel S, et al. (2013) Effect of Acute Exercise on Prostate Cancer Cell Growth. PLoS ONE 8(7): e67579. doi:10.1371/journal.pone.0067579

...Pools of exercise serum from 10 individuals and corresponding rest serum were analyzed with a human protein growth factor array kit (RayBiotech, Norcross, GA, USA). 200 µL of pooled exercise and rest serum was diluted with 1× blocking serum (provided in the kit) while the membranes were blocked for 1 hour in the same blocking buffer. 1 ml of diluted serum was added and incubated at room temperature for 2 hours. The membranes were then analyzed according to the manufacturer's instructions. Chemiluminiscens detection was done on the ImageQuant LAS 4000 (GE Healthcare, Buckinghamshire, UK). Density of individual spots was quantified with Image J software...

Rolfo A, Giuffrida D, Nuzzo AM, Pierobon D, Cardaropoli S, et al. (2013) Pro-Inflammatory Profile of Preeclamptic Placental Mesenchymal Stromal Cells: New Insights into the Etiopathogenesis of Preeclampsia. PLoS ONE 8(3): e59403. doi:10.1371/journal.pone.0059403.

...To investigate differences in cytokines production between normal and preeclamptic PDMSCs, the RayBio Human Cytokine Antibody Array (#AAH-CYT-5, RayBiotech Inc, GA, USA) was used. This specific array was chosen because it allows contemporary detection and quantification of eighty different human cytokines, chemokines and inflammation-related growth factors. Arrays were performed on unconditioned and conditioned culture media from normal and PE PDMSCs (prepared as described above) following manufacturer instructions. Cytokines levels were quantified by densitometric analysis using ImageQuant software and normalized by setting the positive controls as 100 and negative controls as zero percent. The arrays were performed in duplicate on CMs derived from four normal term PDMSCs lines, four preeclamptic PDMSCs lines and on unconditioned media (UCM) as baseline for molecules already present in the culture media. UCM data were used to normalize Normal and PE PDMSCs CM results. Cell lines used were all derived from different control and PE patients...

P-L. Kuo, M-S. Huang, S-K. Huang, W-C. Ni, J-Y. Hung, Y-C. Ko, C-H. Hung, Y-M. Tsai, T-H. Duh and Y-L. Hsu. Signalling pathway of isophorone diisocyanate-responsive interleukin-8 in airway smooth muscle cells. doi10.1183/09031936.00192109 ERJ May 1, 2011 vol. 37 no. 5 1226-1236.

... The profile of cytokines expressed by IPDI-treated BEAS-2B cells was also assessed by RayBio® Human Inflammation Antibody Array (RayBiotech, Inc., Norcross, GA, USA) according to the manufacturer’s instructions...

Malla R, Gopinath S, Alapati K, Gondi CS, Gujrati M, et al. (2010) Downregulation of uPAR and Cathepsin B Induces Apoptosis via Regulation of Bcl-2 and Bax and Inhibition of the PI3K/Akt Pathway in Gliomas. PLoS ONE 5(10): e13731. doi:10.1371/journal.pone.0013731

...Cell lysates from SV- and pCU-treated U251 and 5310 cells were analyzed using a human apoptosis antibody array (RayBiotech, Norcross, GA) according to the manufacturer's instructions. Signal intensities were quantified by densitometry, and fold change was calculated by comparing with controls...

p>Kellar KL, Gehrke J, Weis SE, Mahmutovic-Mayhew A, Davila B, et al. (2011) Multiple Cytokines Are Released When Blood from Patients with Tuberculosis Is Stimulated with Mycobacterium tuberculosis Antigens. PLoS ONE 6(11): e26545. doi:10.1371/journal.pone.0026545.

...A commercial quantitative immuno-microarray (Microarray, Quantibody Human Cytokine Array 1, RayBiotech, Inc., Norcross, GA) was used to measure the concentrations of 20 cytokines in plasma samples; specifically, IFN-γ, IL-1α, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12, IL-13, GM-CSF, GRO, MCP-1, MIP-1α, MIP-1β, MMP-9, RANTES, TNF-α, and VEGF were measured. The manufacturer's procedure was followed and the results were analyzed with the RayBiotech Q Analyzer program. Samples and standards were diluted in the Sample Diluent provided with each kit. The standard curves for each cytokine varied in range from a low of 5–400 pg/mL for IL-13 to 370–30,000 pg/mL for MMP-9. The procedure was similar to that used for the in-house MMIA and ELISA, with the detection antibodies labeled with Alexa 555 that was measured with an Axon Gene Pix 4200 A laser scanner (Molecular Devices, Silicon Valley, CA) at a wavelength of 555 and settings of 450, 550 and 650 pmt. Standard curves were analyzed by regression based on linear and logarithmic transformations of the data. The results of any one assay were based on the same pmt readings and data transformation for all cytokines if the correlation coefficient was ≥0.90 for the standard curve. Data not meeting this latter criterion were excluded...

Simona Giorgini, Daniela Trisciuoglio, Chiara Gabellini, Marianna Desideri, Laura Castellini, Cristina Colarossi, Uwe Zangemeister-Wittke, Gabriella Zupi and Donatella Del Bufalo. Modulation of bcl-xL in Tumor Cells Regulates Angiogenesis through CXCL8 Expression. doi: 10.1158/1541-7786.MCR-07-0088. Mol Cancer Res August 2007 5; 7

...The Human Angiogenesis Antibody Array I (RayBiotech. Inc.) was used according to the manufacturer's protocol to evaluate the secretion of 20 angiogenic factors into the CM of the different lines. A schematic representation of the proangiogenic factors that may be detected by the use of the array has been reported in Fig. 3C. Membranes spotted in duplicate with antibodies against angiogenic factors were blocked with blocking buffer and then were incubated overnight with CM. Next, membranes were washed with wash buffer, incubated with biotin-conjugated antibodies against proangiogenic factors, washed with wash buffer, and incubated with horseradish peroxidase–conjugated streptavidin. The signals on the membranes were detected by chemiluminescence. Membranes, blocking and wash buffer, and antibodies against proangiogenic factors were all provided with the kit. The intensity of protein signal (two spots for each protein) was compared with the relative positive signals by densitometric analysis.

Ying Wang, Dali Cai, Cornelia Brendel, Christine Barett, Philipp Erben, Paul W. Manley, Andreas Hochhaus, Andreas Neubauer and Andreas Burchert. Adaptive secretion of granulocyte-macrophage colony-stimulating factor (GM-CSF) mediates imatinib and nilotinib resistance in BCR/ABL + progenitors via JAK-2/STAT-5 pathway activation. 2007 109: 2147-2155. Prepublished online November 7, 2006; doi:10.1182/blood-2006-08-040022

...A cytokine antibody array was performed using the RayBio Human Cytokine Antibody Array C Series 1000 Kit (Ray Biotech, Norcross, GA) according to the manufacturer’s recommendations. Briefly, 2 membranes each consisting of 60 cytokine antibodies spotted in duplicates onto the membranes were blocked with 2 mL 1 X blocking buffer at room temperature for 30 minutes. Membranes were then incubated with 2 mL serum-free conditioned media derived from LAMA subclones 25UR and 25R, respectively, at room temperature for 90 minutes. Membranes were washed 3 times for 5 minutes at room temperature with 2 mL of 1  wash buffer I, and 2 times with 2 mL of 1 X wash buffer II according to the manufacturer’s recommendations (Ray Biotech). Membranes were then exposed for 90 minutes at room temperature to 1 mL of a 1:500 dilution of biotin-conjugated antibodies. Following a thorough wash as before, membranes were incubated with 2 mL 1:1000 diluted HRP-conjugated streptavidin at room temperature for 120 minutes. Membranes were washed again and exposed for 2 minutes to the peroxidase substrate, which was constituted by mixing 1 X detection buffer C and 1 X Buffer D in a 1:1 ratio. Membranes were exposed to BioMax-MR films (Eastman Kodak, Rochester, NY) for the appropriate times.

Yumi Shiraki, Yoshio Ishibashi, Masataro Hiruma, Akemi Nishikawa and Shigaku Ikeda. Cytokine secretion profiles of human keratinocytes during Trichophyton tonsurans and Arthroderma benhamiae infections. doi: 10.1099/jmm.0.46632-0 J Med Microbiol September 2006 vol. 55 no. 9 1175-1185

...Cytokine release from keratinocytes was analysed by using Human Inflammation Antibody Array III (Ray Biotech) according to the manufacturer's instructions. Briefly, the cytokine array membranes were blocked with 1× blocking buffer for 30 min and then incubated overnight with 1 ml sample at 4 °C. After incubation, the membranes were washed three times with 2 ml 1× Wash Buffer I (Ray Biotech) followed by two washes with 2 ml 1× Wash Buffer II (Ray Biotech) at room temperature with shaking. The membranes were then incubated with 2 ml 1 : 500-diluted biotin-conjugated antibodies for 2 h at room temperature and washed as described above; this was followed by incubation with 1 ml 1 : 40 000-diluted streptavidin-conjugated peroxidase for 1 h at room temperature. After a thorough wash, the membranes were exposed to a peroxidase substrate (detection buffers C and D; Ray Biotech) for 5 min in the dark prior to imaging. The membranes were exposed to an X-ray film within 30 min of exposure to the substrate. Signal intensities were quantified with Scanalyse software (M. Eisen, Lawrence Berkeley National Laboratory; http://www.microarrays.org/software.html). Horseradish peroxidase (HRP)-conjugated antibody served as the positive substrate control at six spots and was also used to identify the membrane orientation. For each spot, the net signal intensity level was determined by subtracting the background signal intensity levels from the total raw signal intensity levels...

Michele Boniotto, William J. Jordan, Joyce Eskdale, Alessandro Tossi, Nikolinka Antcheva, Sergio Crovella, Nancy D. Connell and Grant Gallagher. Human β-Defensin 2 Induces a Vigorous Cytokine Response in Peripheral Blood Mononuclear Cells. doi: 10.1128/AAC.50.4.1433-1441.2006. Antimicrob. Agents Chemother. April 2006 vol. 50 no. 4 1433-1441

...Human protein cytokine arrays were purchased from RayBiotech (Norcross, GA), and supernatants from cell cultures were analyzed according to the manufacturer's instructions. Briefly, cytokine array membranes were first treated with a blocking buffer and then washed and incubated with 1 ml of culture supernatant from either hBD-treated or untreated PBMC cultures. The membranes were then incubated at room temperature for 2 h and washed, and 1 ml of biotin-conjugated detection antibodies was added. Following a further incubation of 2 h at room temperature, the membranes were again washed and finally incubated with 2 ml of streptavidin-horseradish peroxidase at room temperature for 30 min. Finally, the membranes were developed using the kit's enhanced chemiluminescence system, and the results were visualized on X-ray film. Two types of arrays were used. “Cytokine array V” allowed a semiquantitative analysis of the levels of 79 different cytokines or chemokines. “Cytokine array III” allowed the estimation of relative quantities with standard deviations for 42 cytokines...

Lenka Palová-Jelínková, Daniela Rožková, Barbara Pecharová, Jiřina Bártová‡, Anna Šedivá, Helena Tlaskalová-Hogenová, Radek Spíšek and Ludmila Tučková. Gliadin Fragments Induce Phenotypic and Functional Maturation of Human Dendritic Cells. The Journal of Immunology November 15, 2005. vol. 175 no. 10 7038-7045.

...The spectrum of cytokines produced by DC untreated and treated with gliadin fragments (100 μg/ml) was tested using Ab-based protein microarray (RayBio Human Cytokine Ab Array III and 3.1 Map; RayBiotech) designed to detect 42 growth factors, cytokines, or chemokines. Experiments were performed as recommended by manufacturer. Proteins were detected by enhanced chemiluminescence visualized using Fuji Film Las-1000...

Michal Beránek, Petr Kolar, Svatava Tschoplova, Katerina Kankova, Anna Vasku. Genetic variations and plasma levels of gelatinase A (matrix metalloproteinase-2) and gelatinase B (matrix metalloproteinase-9) in proliferative diabetic retinopathy. Molecular Vision 2008; 14:1114-1121 Received 07 November 2007 | Accepted 05 May 2008 | Published 14 June 2008

....... For determination of plasma levels of MMPs the human MMP-2 and MMP-9 ELISA Kits (RayBiotech, Inc., Norcross, GA)  ...