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7982
PathScan® RTK Signaling Antibody Array Kit (Chemiluminescent Readout)
ELISA Kits
Antibody Array Kit

PathScan® RTK Signaling Antibody Array Kit (Chemiluminescent Readout) #7982

Citations (7)
PathScan® RTK Signaling Antibody Array Kit (Chemiluminescent Readout): Image 1

Figure 1. Screening of a panel of cell lines using the PathScan® RTK Signaling Antibody Array Kit (Chemiluminescent Readout) #7982 reveals various phosphorylated RTKs and signaling nodes. A431 cells were starved for 24 hours, then treated with 100 ng/ml EGF #8916 for 3 minutes at 37ºC. NIH/3T3 cells were starved for 24 hours, then treated with 100 ng/ml PDGF #9909 for 5 minutes at 37ºC. Karpas-299 and K562 cells were lysed without starvation or treatment. The array images were captured using chemiluminescent film, with 2-5 second exposure times.

PathScan® RTK Signaling Antibody Array Kit (Chemiluminescent Readout): Image 2

Figure 2. Treatment of MCF-7 cells with IGF-I stimulates phosphorylation of IGF-IR at tyrosine residues, Akt at Ser473 and p44/42 MAPK at Thr202/Tyr204 as detected by the PathScan® RTK Signaling Antibody Array Kit (Chemiluminescent Readout) #7982. MCF-7 cells were starved for 24 hours, then treated with 100 ng/ml IGF-I #3093 for 5 minutes at 37ºC. The chemiluminescent film image (lower panel) and the quantification of that image (upper panel) are shown. The chemiluminescent array images were captured following 2-5 second film exposures.

PathScan® RTK Signaling Antibody Array Kit (Chemiluminescent Readout): Image 3

Figure 3. Treatment of A431 cells with EGF stimulates phosphorylation of EGFR, Akt, p44/42 MAPK and Stat3 as detected by the PathScan® RTK Signaling Antibody Array Kit (Chemiluminescent Readout) #7982. A431 cells were starved for 24 hours and treated with 100 ng/ml EGF #8916 for 5 or 40 minutes. In some cases, cells were treated with either 1 μM wortmannin #9951 for 1 hour before or 1 μM gefitinib for 2 hours before EGF stimulation. Array image pixel intensities obtained from a digital imager are shown in the top panel, while western blots are shown in the bottom panel.

PathScan® RTK Signaling Antibody Array Kit (Chemiluminescent Readout): Image 4

Figure 4. The relationship between lysate protein concentration from untreated and IGF-I treated MCF-7 cells and the pixel intensities of phospho-IGF-IR (pan-Tyr), phospho-Akt (Ser473) and phospho-p44/42 (Thr202/Tyr204) is shown. MCF-7 cells were starved for 24 hours, then treated with 100 ng/ml IGF-I #3093 for 5 minutes at 37ºC.

PathScan® RTK Signaling Antibody Array Kit (Chemiluminescent Readout): Image 5

Figure 5. Target map of the PathScan® RTK Signaling Antibody Array Kit (Chemiluminscent Readout)

To Purchase # 7982

Product Description

The PathScan® RTK Signaling Antibody Array Kit (Chemiluminescent Readout) is a slide-based antibody array founded upon the sandwich immunoassay principle. The array kit allows for the simultaneous detection of 28 receptor tyrosine kinases and 11 important signaling nodes, when phosphorylated at tyrosine or other residues. Target-specific capture antibodies have been spotted in duplicate onto nitrocellulose-coated glass slides. Each kit contains two 8-pad slides, allowing the user to test up to 16 samples. Cell lysate is incubated on the slide followed by a biotinylated detection antibody cocktail. Streptavidin-conjugated HRP and LumiGLO® Reagent are then used to visualize the bound detection antibody by chemiluminescence. An image of the slide can be captured with either a digital imaging system or standard chemiluminescent film. The image can be analyzed visually or the 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

Cell Signaling Technology's PathScan® RTK Signaling Antibody Array Kit detects the indicated RTKs and signaling nodes only when phosphorylated at tyrosine or specified residues (see Array Target Map). No significant crossreactivity has been observed between targets, with the exception of some crossreactivity of the FLT3 antibody with phosphorylated EphB3. In addition, Stat1 (Tyr701) and Stat3 (Tyr705) may be detected when phosphorylated at other tyrosine sites within the proteins. This kit is optimized for cell lysates diluted to a total protein concentration between 0.2 and 1 mg/ml (see Figure 4). All capture antibodies have been validated for human targets. Although this kit has not been tested with mouse lysates, it is expected than many capture antibodies will crossreact in murine systems.

Background

Receptor Tyrosine Kinases (RTKs) are a family of cell surface receptors that signal primarily through tyrosine phosphorylation events (1). RTKs trigger a wide range of downstream signaling cascades, including the PI3K/Akt, MAPK and Jak/Stat pathways. These pathways control basic cellular functions such as division, growth, metabolism, differentiation, migration and survival. Dysregulation of RTK signaling has been implicated in a large number of cancers (2), making RTKs popular targets for pharmaceutical intervention.

  1. Schlessinger, J. (2000) Cell 103, 211-25.
  2. Blume-Jensen, P. and Hunter, T. (2001) Nature 411, 355-65.

Pathways & Proteins

Explore pathways + proteins related to this product.

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.
LumiGLO is a registered trademark of Kirkegaard & Perry Laboratories.