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88334
CellSimple™ Akt and MAPK Activation Antibody Assay Kit
Assay Kit

CellSimple Akt and MAPK Activation Antibody Assay Kit #88334

Citations (0)
CellSimple cell-based analysis of Jurkat cells using the CellSimple Akt and MAPK Activation Antibody Assay Kit. Cells were subjected to the following treatments: A) LY294002 #9901 (10 μM), Wortmannin #9951 (1 μM), and U0126 #9903 (2 μM) for 2.5 hr; B) U0126 (2 μM, 2.5 hr); C) LY294002 (10 μM) and Wortmannin (1 μM, 2.5 hr) followed by TPA #4174 (1 μM, 15 min); D) No addition of any kinase inhibitors followed by treatment of cells with TPA (1 μM, 15 min).

Fixed and permeabilized cells were stained and analyzed using the Open Flow Cytometry application. Mean fluorescence intensity (MFI) of Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate) in the green channel

(525/45 nm filter; x-axis) and Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (197G2) Rabbit mAb (PE Conjugate) in the red channel (561 nm/LP filter; y-axis) is shown in each dot plot. Cell concentration, MFIs, and percentages are displayed in the boxes adjacent to each quadrant. Instrument screen shots are shown.

Inquiry Info.# 88334

Product Description

The CellSimple Akt and MAPK Activation Antibody Assay Kit is designed for use with the CellSimple Cell Analyzer. The kit contains Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (197G2) Rabbit mAb (PE Conjugate) #14095 and Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate) #4071. The kit allows for the quick and easy measurement of relative phosphorylation levels of ERK 1/2 MAPK and Akt under given experimental conditions.

Specificity / Sensitivity

Antibodies provided in the CellSimple  Akt and MAPK Activation Antibody Assay Kit detect endogenous levels of their respective target. Additionally the Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (197G2) Rabbit mAb (PE Conjugate) does not cross-react with the corresponding phosphorylated residues of either JNK/SAPK or p38 MAP kinase.

Species Reactivity:

Human, Mouse

Background

The CellSimple Cell Analyzer is a benchtop instrument that utilizes a disposable thin-film cassette and a combination of a 488 nm laser, two photomultiplier tubes (525/45 nm and 561 nm LP filters), Coulter Principle-based cell measurements, and on-board software to provide easy-to-run applications and data analysis. Data acquisition occurs within approximately 10 seconds per test. The instrument relies on disposable cassettes for sample handling, which alleviates the need for flow cell cleaning and fluidics maintenance and the instrument is small enough to be portable between the lab bench and the hood. Applications include quantitative assessments of cell viability, apoptosis, other labeled antibody markers and single and multiplexed bead-based assays for protein and cellular analysis.
The MAP Kinase (MAPK) and the PI-3 Kinase/Akt pathways are two of the most well characterized and widely studied signaling modules which transmit signals emanating from the extracellular environment to alter cell behavior. They are involved in a broad range of cellular processes such as cell proliferation, survival, differentiation, and motility. The ERK 1 and 2 (from the MAPK pathway) and Akt protein kinases are two quintessential components serving as integration hubs in their respective pathways. ERK 1/2 protein kinases are activated by dual phosphorylation on Thr202 and Tyr204 from the the dual specificity kinases MEK1 and MEK2. The Akt protein kinase is activated by phosphorylation on Ser473 and Thr308 from the TORC2 complex and PDK1, respectively. The relative levels of these phosphorylations are reliable proxies of activation levels of these kinases. The knowledge about the levels of activation of these kinases is instructive in a broad range of experimental settings making these two molecular events some of the most popular and sought after assay endpoints.

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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This product is provided under an intellectual property license from Life Technologies Corporation. The transfer of this product is conditioned on the buyer using the purchased product solely in research conducted by the buyer, excluding contract research or any fee for service research, and the buyer must not (1) use this product or its components for (a) diagnostic, therapeutic or prophylactic purposes; (b) testing, analysis or screening services, or information in return for compensation on a per-test basis; or (c) manufacturing or quality assurance or quality control, and/or (2) sell or transfer this product or its components for resale, whether or not resold for use in research. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or [email protected].
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