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Human Cytokine Array G5

Reactivity: Human AA Fluorometric Semi-Quantitative Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Lysate Glass Slide
Catalog No. ABIN625585
  • Reactivity
    Human
    Detection Method
    Fluorometric
    Method Type
    Sandwich ELISA
    Application
    Antibody Array (AA)
    Purpose
    G-Series Human Cytokine Antibody Array 5 Kit. Detects 80 Human Cytokines. Suitable for all liquid sample types.
    Brand
    RayBio®
    Sample Type
    Serum, Plasma, Cell Culture Supernatant, Cell Lysate, Tissue Lysate
    Analytical Method
    Semi-Quantitative
    Specificity
    ENA-78 (CXCL5), GCSF, GM-CSF, GRO alpha/beta/gamma, GRO alpha (CXCL1), I-309 (TCA-3/CCL1), IL-1 alpha (IL-1 F1), IL-1 beta (IL-1 F2), IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8 (CXCL8), IL-10, IL-12 p40/p70, IL-13, IL-15, IFN-gamma, MCP-1 (CCL2), MCP-2 (CCL8), MCP-3 (MARC/CCL7), M-CSF, MDC (CCL22), MIG (CXCL9), MIP-1 beta (CCL4), MIP-1 delta (CCL15), RANTES (CCL5), SCF, SDF-1 alpha (CXCL12 alpha), TARC (CCL17), TGF beta 1, TNF alpha, TNF beta (TNFSF1B), EGF, IGF-1, Angiogenin, Oncostatin M, Thrombopoietin (TPO), VEGF-A, PDGF-BB, Leptin, BDNF, BLC (CXCL13), Ck beta 8-1 (CCL23), Eotaxin-1 (CCL11), Eotaxin-2 (MPIF-2/CCL24), Eotaxin-3 (CCL26), FGF-4, FGF-6, FGF-7 (KGF), FGF-9, Flt-3 Ligand, Fractalkine (CX3CL1), GCP-2 (CXCL6), GDNF, HGF, IGFBP-1, IGFBP-2, IGFBP-3, IGFBP-4, IL-16, IP-10 (CXCL10), LIF, Light (TNFSF14), MCP-4 (CCL13), MIF, MIP-3 alpha (CCL20), NAP-2 (PPBP/CXCL7), NT-3, NT-4, Osteopontin (SPP1), Osteoprotegerin (TNFRSF11B), PARC (CCL18), PLGF, TGF beta 2, TGF beta 3, TIMP-1, TIMP-2
    Characteristics
    • High sensitivity and specificity
    • Low sample volume (10-100 μL per array)
    • Large dynamic range of detection
    • Compatible with most sample types
    • Test 4 or 8 samples on each slide
    • Suitable for high-throughput assays
    Components
    Cytokine Antibody Array glass slide (4 or 8 arrays per slide)
    Biotinylated Detection Antibodies
    Streptavidin-conjugated HiLytePlus™ 555 Fluor
    Blocking Buffer
    20X Wash Buffer I
    20X Wash Buffer II
    2X Cell Lysis Buffer
    G-Series Antibody Array accessories
    Accessories include: 16-well incubation chamber with gasket, protective cover, snap-on sides, adhesive film
    Material not included
    Small plastic boxes or containers
    Pipettors, pipette tips and other common lab consumables
    Orbital shaker or oscillating rocker
    Aluminum foil
    Gene microarray scanner or similar laser fluorescence scanner
  • Application Notes
    Completely cover array area with sample or buffer during incubation. Avoid foaming during incubation steps. Perform all incubation and wash steps under gentle rocking or rotation. Cover the incubation chamber with adhesive film during incubation, particularly when incubation is more than 2 hours or <70 μL of sample or reagent is used. Several incubation steps such as step 6 (blocking), step 7 (sample incubation), step 10 (detection antibody incubation), or step 13 (Cy3 equivalent dyestreptavidin incubation) may be done overnight at 4 °C. Please make sure to cover the incubation chamber tightly to prevent evaporation.
    Comment

    The G-Series arrays feature fluorescent signal detection. The antibodies are spotted on glass slide solid supports and require a laser scanner for data collection.
    All G-Series arrays work on the sandwich ELISA principle, utilizing a matched pair of antibodies: an immobilized capture antibody and a corresponding biotinylated detection antibody.

    Sample Volume
    100 μL
    Assay Time
    6 h
    Plate
    Glass Slide
    Protocol
    1. Dry the glass slide
    2. Block array surface
    3. Incubate with Sample
    4. Incubate with Biotinylated Detection Antibody Cocktail
    5. Incubate with Streptavidin-Conjugated Fluor
    6. Disassemble the glass slide
    7. Scan with a gene microarray laser scanner
    8. Perform densitometry and analysis
    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 µl of original or diluted serum, plasma, cell culture media, or other body fluid, or 50-500 µg/ml of protein for cell and tissue lysates. If you experience high background or if the fluorescent signal intensities exceed the detection range, further dilution of your sample is recommended.

    Assay Procedure

    Take out the glass slide 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 for another 1-2 hours.

    Blocking & Incubation
    1. Add 100 µl Sample Diluent into each well and incubate at room temperature for 30 minutes to block slides.
    2. Decant buffer from each well. Add 100 µl of sample to each well. Incubate arrays at room temperature for 1-2 hour.
    3. Decant the samples from each well, and wash 5 times (5 min each) with 150 µl of 1X Wash Buffer I at room temperature with gentle shaking. Completely remove wash buffer in each wash step. Dilute 20x Wash Buffer I with H2O.
    4. Decant the 1x Wash Buffer I from each well, wash 2 times (5 min each) with 150 µl of 1X Wash Buffer II at room temperature with gentle shaking.Completely remove wash buffer in each wash step. Dilute 20X Wash Buffer II with H2O.

    Incubation with Biotinylated Antibody Cocktail & Wash
    5. Reconstitute the detection antibody by adding 1.4 ml of Sample Diluent to the tube. Spin briefly.
    6. Add 80 µl of the detection antibody cocktail to each well. Incubate at room temperature for 1-2 hour.
    7. Decant the samples from each well, and wash 5 times (5 mins each) with 150 µl of 1X Wash Buffer I and then 2 times with 150 µl of 1x Wash Buffer II at room temperature with gentle shaking. Completely remove wash buffer in each wash step.

    Incubation with Cy3 Equivalent Dye-Streptavidin & Wash
    8. After briefly spinning down, add 1.4 ml of Sample Diluent to Cy3 equivalent dye-conjugated streptavidin tube. Mix gently.
    9. Add 80 µl of Cy3 equivalent dye-conjugated streptavidin to each well. Cover the device with aluminum foil to avoid exposure to light or incubate in dark room. Incubate at room temperature for 1 hour.
    10. Decant the samples from each well, and wash 5 times (5 mins each) with 150 µl of 1X Wash Buffer I at room temperature with gentle shaking. Completely remove wash buffer in each wash step.

    Fluorescence Detection
    11. Disassemble the device by pushing clips outward from the slide side. Carefully remove the slide from the gasket.
    12. Place the slide in the Slide Washer/Dryer (a 4-slide holder/centrifuge tube), add enough 1x Wash Buffer I (about 30 ml) to cover the whole slide, and then gently shake at room temperature for 15 minutes. Decant Wash Buffer I. Wash with 1x Wash Buffer II (about 30 ml) and gently shake at room temperature for 5 minutes.
    13. Remove water droplets completely by gently applying suction with a pipette to remove water droplets. Do not touch the array, only the sides.
    14. Imaging: The signals can be visualized through use of a laser scanner equipped with a Cy3 wavelength (green channel) such as Axon GenePix.

    Calculation of Results

    Data extraction can be done using the GAL file that is specific for this array along with the microarray analysis software (GenePix, ScanArray Express, ArrayVision, MicroVigene, etc.).

    Restrictions
    For Research Use only
  • Handling Advice
    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 powder free gloves. Handle glass slide/s in clean environment. The G-Series slides do not have bar codes. To help distinguish one slide from another, transcribe the slide serial number from the slide bag to the back of the slide with a fine point permanent marker. Please write the number on the very bottom edge of the slide, taking care to avoid writing on the array well areas.
    Storage
    -20 °C
    Storage Comment
    For best results, store the entire kit frozen at -20°C upon arrival. Stored frozen, the kit will be stable for at least 6 months which is the duration of the product warranty period. Once thawed, store array slide(s) and 1X Blocking Buffer at -20°C and all other reagents undiluted at 4°C for no more than 3 months.
    Expiry Date
    6 months
  • Ma, Kua, Lim, Lee, Chua: "In vitro characterization of human hair follicle dermal sheath mesenchymal stromal cells and their potential in enhancing diabetic wound healing." in: Cytotherapy, Vol. 17, Issue 8, pp. 1036-51, (2016) (PubMed).

    Wolf, Siegert, Rothmiller, Scheithauer, Strobelt, Steinritz, Worek, Thiermann, Schmidt: "Characterization of sulfur mustard resistant keratinocyte cell line HaCaT/SM." in: Toxicology letters, Vol. 244, pp. 49-55, (2016) (PubMed).

    Chen, Shi, Feng, Kong, Chen, Geng, Chen, Liu, Li, Chen, Gao, Sun: "Leptin and Neutrophil-Activating Peptide 2 Promote Mesenchymal Stem Cell Senescence Through Activation of the Phosphatidylinositol 3-Kinase/Akt Pathway in Patients With Systemic Lupus Erythematosus." in: Arthritis & rheumatology (Hoboken, N.J.), Vol. 67, Issue 9, pp. 2383-93, (2015) (PubMed).

    Zhong, Zhou, Wu, Guo, Tan, Zhang, Zhang, Geng, Pan, Luo, Zhang, Xu, Liu, Liu, Gao, Liu, Ren, Li, Zhou, Zhang: "A SnoRNA-derived piRNA interacts with human interleukin-4 pre-mRNA and induces its decay in nuclear exosomes." in: Nucleic acids research, Vol. 43, Issue 21, pp. 10474-91, (2015) (PubMed).

    Chollet-Hinton, Stuebe, Casbas-Hernandez, Chetwynd, Troester: "Temporal Trends in the Inflammatory Cytokine Profile of Human Breastmilk." in: Breastfeeding medicine : the official journal of the Academy of Breastfeeding Medicine, (2014) (PubMed).

    Zhen, Krutzik, Levin, Kasparian, Zack, Kitchen: "CD4 ligation on human blood monocytes triggers macrophage differentiation and enhances HIV infection." in: Journal of virology, Vol. 88, Issue 17, pp. 9934-46, (2014) (PubMed).

    Rath, Fair, Jamal, Camphausen, Tofilon: "Astrocytes enhance the invasion potential of glioblastoma stem-like cells." in: PLoS ONE, Vol. 8, Issue 1, pp. e54752, (2013) (PubMed).

    Choi, Kim, Yoon, Cho: "Full-thickness skin wound healing using human placenta-derived extracellular matrix containing bioactive molecules." in: Tissue engineering. Part A, Vol. 19, Issue 3-4, pp. 329-39, (2013) (PubMed).

    Stewart, Yang, Makowski, Troester: "Basal-like breast cancer cells induce phenotypic and genomic changes in macrophages." in: Molecular cancer research : MCR, Vol. 10, Issue 6, pp. 727-38, (2012) (PubMed).

    Taxman, Holley-Guthrie, Huang, Moore, Bergstralh, Allen, Lei, Gris, Ting: "The NLR adaptor ASC/PYCARD regulates DUSP10, mitogen-activated protein kinase (MAPK), and chemokine induction independent of the inflammasome." in: The Journal of biological chemistry, Vol. 286, Issue 22, pp. 19605-16, (2011) (PubMed).

    Smalley-Freed, Efimov, Short, Jia, Zhao, Washington, Robine, Coffey, Reynolds: "Adenoma formation following limited ablation of p120-catenin in the mouse intestine." in: PLoS ONE, Vol. 6, Issue 5, pp. e19880, (2011) (PubMed).

    Li, Sidell: "Growth-related oncogene produced in human breast cancer cells and regulated by Syk protein-tyrosine kinase." in: International journal of cancer. Journal international du cancer, Vol. 117, Issue 1, pp. 14-20, (2005) (PubMed).

  • 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.
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