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PathScan® Signaling Nodes II 4-Plex Array Kit

PathScan^® Signaling Nodes II 4-Plex Array Kit #7139

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Product Information

Product Description

CST’s PathScan^® Signaling Nodes II 4-Plex Array Kit is based upon the sandwich ELISA principle. Rather than one immobilized antibody, these PathScan^® 4-Plex Array Kits contain four different antibodies in each well of a 96-well plate. This allows the researcher to measure four different target proteins from a single well simultaneously. In addition, the incorporation of multiple phospho-specific antibody pairs allows the researcher to measure the activation state of important signaling nodes. The PathScan^® Signaling Nodes II 4-Plex Array Kit allows the detection of: Phospho-Akt1 (Ser473); Phospho-MEK1 (Ser217/221); Phospho-p44 MAPK (Thr202/Tyr204); and Total Akt1.
Briefly, 4 capture antibodies with distinct target specificity have been spotted onto the bottom of each well of a microwell plate. After incubation with cell lysates, the spotted antibodies capture the target proteins. Following extensive washing, a mixture of detection antibodies is added to detect the captured target proteins. An HRP-linked secondary solution is then used to recognize the bound detection antibodies. Chemiluminescent HRP substrate is used to produce luminescent signal.

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^® Signaling Nodes II 4-Plex Array Kit detects endogenous levels of all target proteins. As shown in Figure 2, stimulation of NIH/3T3 cells with PDGF promotes phosphorylation of Akt at Ser473, p44 MAP kinase at Thr202/Tyr204 and MEK1 on Ser217/221. Levels of total Akt1 remain unchanged.

Background

Cells must respond in an appropriate fashion to many complex signaling events. Extracellular signaling cues are organized into well defined signal transduction modules that control fundamental cellular behavior. Two prominent signaling modules that are among the best characterized are the p44/42 MAP kinase (ERK MAPK) and Akt signal transduction pathways. These signaling modules control cellular growth, proliferation, movement, survival, and death. The p44/42 MAPK is activated by a wide variety of extracellular signals including growth and neurotrophic factors, cytokiones, hormones, and neurotransmitters. p44/42 MAPK activation occurs through phosphorylation of theronine and tyrosine at the sequence T*EY* by a dual specificity kinase called MAP kinase kinase (MEK). The Akt protein kinase is generally activated in response to growth factor stimulation and phosphorylated on Ser473. Growth factor stimulation results in phosphorylation of S6 Ribosomal protein on Ser235/236, leading to an increase in protein synthesis and cell cycle progression. Phosphorylation levels of critical molecular switches such as MAPKs and Akt therefore serve as a reliable indicator of the activation state of the entire signaling module. The profiling of phosphorlation events using phospho-specific antibodies is now widely used to investigate diagnostic pathology (1,2). The PathScan^® Signaling Nodes II 4-Plex Array Kit provides the researcher with means to profile numerous chemical compounds and obtain in-cell relative potency (3).

Database Links

UniProt ID: P27361 , P36507 , P31751 , Q9Y243 , P28482 , P31749 , Q02750

Entrez-Gene Id: 5595 , 5605 , 208 , 10000 , 5594 , 207 , 5604

Limited Uses

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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 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 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.

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PathScan is a registered trademark of Cell Signaling Technology, Inc.

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PathScan® Signaling Nodes II 4-Plex Array Kit #7139