Human Cytokine Array 5

Details for Product No. ABIN625729
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Reactivity
Human
Application
Antibody Array (AA), Multiplex ELISA
Pubmed 4 references available
Quantity 1 array
Quantity Comment 32-50 Samples
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Catalog No. ABIN625729
5,052.30 $
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Brand Quantibody®
Specificity Quantibody® Human Cytokine Array 5 detects: Adiponectin, Adipsin, AFP, ANGPTL4, B2M, BCAM, CA125, CA15-3, CEA, CRP, ErbB2, Follistatin, FSH, GROa, HCGb, IGF-1 SR, IL-1 sRII, IL-3, IL-18 Rb, IL-21, Leptin, MMP-1, MMP-2, MMP-3, MMP-8, MMP-9, MMP-10, MMP-13,
Sensitivity Fluorescence with laser scanner: Cy3 equivalent dye
Characteristics Quantibody® Human Cytokine Array 5, multiplex ELISA array Kit for quantitative measurement of 40 Cytokines in 8-10 samples. Suitable for all sample Types
Components 1. Quantibody Array Glass Chip (1 in 1-Slide Kit, 2 in 2-Slide Kit) 2. Sample Diluent (1 each) 3. 20X Wash Buffer I (2 in 1-Slide Kit, 3 in 2-Slide Kit) 4. 20X Wash Buffer II (1 each) 5. Lyophilized cytokine standard mix * (1 each) 6. Detection antibody cocktail (1 in 1-Slide Kit, 2 in 2-Slide Kit) 7. Cy3 equivalent dye-conjugated Streptavidin (1 in 1-Slide Kit, 2 in 2-Slide Kit) 8. Slide Washer/Dryer (1 each) 9. Adhesive device sealer (5 in 1-Slide Kit, 10 in 2-Slide Kit) 10. Manual 1 1 * See Section VI for detailed cytokine concentrations after reconstitution
Material not included Orbital shaker. Laser scanner for fluorescence detection. Aluminum foil Distilled water. 1.5ml Polypropylene microcentrifuge tubes
Background Cytokines play an important role in innate immunity, apoptosis, angiogenesis, cell growth and differentiation. They are involved in interactions between different cell types, cellular responses to environmental conditions, and maintenance of homeostasis. In addition, cytokines are also involved in most disease processes, including cancer and cardiac diseases. The traditional method for cytokine detection and quantification is through the use of an enzyme-linked immunosorbent array (ELISA). In this method, target protein is first immobilized to a solid support. The immobilized protein is then complexed with an antibody that is linked to an enzyme. Detection of the enzyme-complex can then be visualized through the use of a substrate that produces a detectable signal. While the traditional method works well for a single protein, the overall procedure is time consuming and requires a lot of sample. With little sample to work with, conservation of precious small quantities becomes a risky task. Take the advantage of advancement in microarray technology over the last decade, more and more choices are available to the scientist today. A long-standing leader in the field, Raybiotech, has pioneered the development of cytokine antibody arrays, which has now been widely applied in the research community with hundreds of peer reviewed publications such as in Cell and Nature. Quantibody® array, our quantitative array platform, uses the multiplexed sandwich ELISA-based technology and enables researchers to accurately determine the concentration of multiple cytokines simultaneously. It combines the advantages of the high detection sensitivity / specificity of ELISA and the high throughput of the arrays. Like a traditional sandwich- based ELISA, it uses a pair of cytokine specific antibodies for detection. A capture antibody is first bound to the glass surface. After incubation with the sample, the target cytokine is trapped on the solid surface. A second biotin- labeled detection antibody is then added, which can recognize a different isotope of the target cytokine. The cytokine-antibody-biotin complex can then be visualized through the addition of the streptavidin-labeled Cy3 equivalent dye using a laser scanner. Unlike the traditional ELISA, Quantibody products use array format. By arraying multiple cytokine specific capture antibodies onto a glass support, multiplex detection of cytokines in one experiment is made possible. In detail, one standard glass slide is spotted with 16 wells of identical cytokine antibody arrays. Each antibody, together with the positive controls is arrayed in quadruplicate. The slide comes with a 16-well removable gasket which allows for the process of 16 samples in one slide. Four slide chips can be nested into a tray, which matches a standard microplate and allows for automated robotic high throughput process of 64 arrays simultaneously. For cytokine quantification, the array specific cytokine standards, whose concentration has been predetermined, are provided to generate a standard curve for each cytokine. In a real experiment, standard cytokines and samples will be assayed in each array simultaneously through a sandwich ELISA procedure. By comparing signals from unknown samples to the standard curve, the cytokine concentration in the samples will be determined. Quantibody® array kits have been confirmed to have similar detection sensitivity as traditional ELISA. Our current high density Quantibody kits allow scientists to quantitatively determine the concentration of 240 human or 120 mouse cytokines in a single experiment. This is not only one of the most efficient products on the market for cytokine quantification, but makes it more affordable for quantification of large number of proteins. Simultaneous detection of multiple cytokines undoubtedly provides a powerful tool for drug and biomarker discovery.
Application Notes 1. Take out the glass chip from the box, and let it equilibrate to room temperature inside the sealed plastic bag for 20-30 minutes. Remove slide from the plastic bag, peel off the cover film, and let it air dry at room temperature for another 1-2 hours. Note: Incomplete drying of slides before use may cause the formation of comet tails. Do not touch the surface of the slides, as the microarray slides are very sensitive. Hold the slides by the edges only. Handle all buffers and slides with latex free gloves. Handle glass chip in clean environment. Because there is no barcode on the slide, transcribe the slide serial number from the slide bag to the back of the slide with a permanent marker before discarding the slide bag. Once the slide is disassembled, you might not have enough info to distinguish one slide from the other.
Comment

Array support Samples Incubation of Sample With arrayed antibody 1-2 hr Supports Cocktail of Biotin-Ab Incubation with 1-2 hr Biotinylated Ab Labeled ­ streptavidin Incubation with 1 hr Cy3 equivalent dye Labeled- streptavidin Detection of signals Data analysis and graph.

Sample Preparation Use serum-free conditioned media if possible. If serum-containing conditioned media is required, it is highly recommended that complete medium be used as a control since many types of sera contains cytokines. We recommend the following parameters for your samples: 50 to 100 l of original or diluted serum, plasma, cell culture media, or other body fluid, or 50-500 g/ml of protein for cell and tissue lysates. If you experience high background or the readings exceed the detection range, further dilution of your sample is recommended.
Assay Procedure Reproducibility: CV < 20%. Assay duration: 6 hrs
Restrictions For Research Use only
Storage -20 °C
Product cited in: Akhtar, Haqqi: "Epigallocatechin-3-gallate suppresses the global interleukin-1beta-induced inflammatory response in human chondrocytes." in: Arthritis research & therapy, Vol. 13, Issue 3, pp. R93, 2011 (PubMed).

Cao, Ren, Gong et al.: "Bronchial epithelial cells release IL-6, CXCL1 and CXCL8 upon mast cell interaction." in: Cytokine, 2011 (PubMed).

Tung, Fraser, Nataatmadja et al.: "Age of blood and recipient factors determine the severity of transfusion-related acute lung injury (TRALI)." in: Critical care (London, England), Vol. 16, Issue 1, pp. R19, 2012 (PubMed).

Li, Wang, Zhang et al.: "Mesenchymal stem cells protect podocytes from apoptosis induced by high glucose via secretion of epithelial growth factor." in: Stem cell research & therapy, Vol. 4, Issue 5, pp. 103, 2013 (PubMed).

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