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13124
PathScan® Th1/Th2/Th17 Cytokine Antibody Array Kit (Fluorescent Readout)
ELISA Kits
Antibody Array Kit

PathScan® Th1/Th2/Th17 Cytokine Antibody Array Kit (Fluorescent Readout) #13124

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PathScan® Th1/Th2/Th17 Cytokine Antibody Array Kit (Fluorescent Readout): Image 1

Figure 1. Target map of the PathScan® Th1/Th2/Th17 Cytokine Antibody Array Kit (Fluorescent Readout) #13124.

PathScan® Th1/Th2/Th17 Cytokine Antibody Array Kit (Fluorescent Readout): Image 2

Figure 2. Human peripheral blood mononuclear cells (PBMCs) were isolated from whole blood and stimulated with immobilized anti-human CD3 antibody, soluble anti-human CD28 antibody (1 μg/ml), hIL-2 #8907 (2 ng/ml) and hIL-4 #8919 (1 ng/ml) for 2 days. Cells were washed and cultured for 3 days with hIL-2 (2 ng/ml) and hIL-4 (1 ng/ml). Cells were then washed and stimulated with TPA #4174 (10 ng/ml) and Ionomycin, Calcium Salt #9995 (1 μg/ml) for 24 hr before supernatant collection. Supernatants were prepared and analyzed using PathScan® Th1/Th2/Th17 Cytokine Antibody Array Kit (Fluorescent Readout) #13124. Panel A shows images that were acquired using LI-COR® Biosciences Odyssey® imaging system. Panels B and C show high and low end raw values of quantified fluorescence intensity. Pixel intensity was quantified using the LI-COR® Image Studio v2.0 array analysis software.

PathScan® Th1/Th2/Th17 Cytokine Antibody Array Kit (Fluorescent Readout): Image 3

Figure 3. Human PBMCs were isolated from whole blood and pre-treated with hIFN-γ #8901 (10 ng/ml) for 2 hr. LPS was then added (1 μg/ml) and incubated for an additional 22 hr before supernatant collection. Supernatants were prepared and analyzed using the PathScan® Th1/Th2/Th17 Cytokine Antibody Array Kit (Fluorescent Readout) #13124. Panel A shows images that were acquired using the LI-COR® Biosciences Odyssey® imaging system. Panel B shows raw values of quantified fluorescence intensity. Pixel intensity was quantified using the LI-COR® Image Studio v2.0 array analysis software.

PathScan® Th1/Th2/Th17 Cytokine Antibody Array Kit (Fluorescent Readout): Image 4

Figure 4. Human PBMCs were isolated from whole blood and CD4+ T cells separated by positive selection. The CD4+ T cells were stimulated with immobilized anti-human CD3 antibody, soluble anti-human CD28 antibody (1 μg/ml), hIL-23 (10 ng/ml), hTGF-β1 #8915 (1 ng/ml), hIL-6 #8904 (20 ng/ml), hTNF-α #8902 (10 ng/ml), hIL-1β #8900 (10 ng/ml), anti-human IL-4 antibody (10 μg/ml) and anti-human IFN-γ antibody (1 μg/ml) for 3 days before supernatant collection. Supernatants were prepared and analyzed using PathScan® Th1/Th2/Th17 Cytokine Antibody Array Kit (Fluorescent Readout) #13124. Panel A shows images that were acquired using LI-COR® Biosciences Odyssey® imaging system. Panels B and C show high and low end raw values of quantified fluorescence intensity. Pixel intensity was quantified using the LI-COR® Image Studio v2.0 array analysis software.

To Purchase # 13124

Product Description

Description: The PathScan® Th1/Th2/Th17 Cytokine Antibody Array Kit (Fluorescent Readout) uses glass slides as the planar surface and is based upon the sandwich immunoassay principle. This array kit allows for the simultaneous detection of 12 unique extracellular signaling molecules. Target-specific capture antibodies have been spotted in duplicate onto nitrocellulose-coated glass slides. Each kit contains two slides allowing the user to test up to 32 different samples and generate 384 data points in a single experiment. Cell supernatant is incubated on the slide followed by a biotinylated detection antibody cocktail. Streptavidin-conjugated Dylight™ 680 is then used to visualize the bound detection antibody. A fluorescent image of the slide can then be captured with a digital imaging system and spot intensities quantified using array analysis software.

Storage

Kit should be stored at 4°C with the exception of Lysis Buffer, which is stored at –20°C (packaged separately).

Specificity / Sensitivity

PathScan® Th1/Th2/Th17 Cytokine Antibody Array Kit (Fluorescent Readout) detects the target proteins as specified on the Array Target Map (figure 1). No significant cross reactivity has been observed between targets. This kit is optimized for cell culture supernatants. Recommended starting cell culture supernatant range is 20-75 μl. All antibodies have been validated for human-derived cell culture supernatants.

Background

Cytokines are secreted intercellular signaling molecules that regulate many biological processes including inflammation, host defense, and cell differentiation. Cytokine profiles may provide insight into the molecular mechanisms that distinguish between healthy and diseased states. The PathScan® Th1/Th2/Th17 Cytokine Antibody Array Kit offers an antibody panel against a broad array of cytokines to enable measurement of their relative changes in cell culture supernatants.

Upon activation, naive CD4+ helper T cells differentiate into distinct functional subsets. The development of these subsets is driven, in part, by the cytokine milieu. Type 1 (Th1) cells help drive cellular immunity against intracellular pathogens. IL-12 and IFN-γ induce Th1 cell development. Th1 cells produce IFN-γ and IL-2, which provide a positive feedback loop to enhance Th1 cell differentiation and NK cell and CD8+ T cell cytolytic activity.

Th2 cells play a crucial role in the humoral immune response against extracellular pathogens. IL-4 drives development of Th2 cells, which subsequently produce IL-4, IL-5, and IL-13. These cytokines induce B cell proliferation, antibody production, IgE class switching, and activate eosinophils respectively.

Another distinct helper T cell lineage, Th17, is important for mucosal immunity. Dysregulation of Th17 may significantly contribute to the development of autoimmunity. IL-17 produced by Th17 cells induce secretion of pro-inflammatory cytokines IL-6, IL-8, GM-CSF, and TNF-α. Many of these molecules link innate and adaptive immunity through the recruitment and activation of innate immune cells.

Effective immune responses require finely tuned coordination between pro and anti-inflammatory signals. Pro-inflammatory molecules play important roles in activating key immune players to fight infection. IL-8 induces granulocyte migration and activates neutrophil phagocytic activity. GM-CSF mobilizes monocytes into infected tissue and activates macrophage and neutrophils. TNF-α is a multifunctional pro-inflammatory cytokine involved with a number of processes including cell proliferation, differentiation, and apoptosis.

Uncontrolled inflammation may damage surrounding host tissue. IL-10 is a prototypical anti-inflammatory cytokine that serves to terminate the acute inflammatory response by inhibiting Th1 cell function and pro-inflammatory cytokine production.

  1. Romagnani, S. (1999) Inflamm Bowel Dis 5, 285-94.
  2. Bradley, L.M. et al. (2000) Immunol Res 21, 149-58.
  3. O'Garra, A. and Arai, N. (2000) Trends Cell Biol 10, 542-50.
  4. Harrington, L.E. et al. (2005) Nat Immunol 6, 1123-32.
  5. Stockinger, B. and Veldhoen, M. (2007) Curr Opin Immunol 19, 281-6.
  6. Köhidai, L. and Csaba, G. (1998) Cytokine 10, 481-6.
  7. Carey, A.J. et al. (2012) JAKSTAT 1, 159-167.

Pathways & Proteins

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Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST's products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST's Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
PathScan is a trademark of Cell Signaling Technology, Inc.
DyLight is a trademark of Thermo Fisher Scientific, Inc. and its subsidiaries.
LI-COR is a registered trademark of LI-COR, Inc.
Odyssey is a registered trademark of LI-COR, Inc.