Cellomics HCS Reagent Kit for activated (phosphorylated) p38 MAPK.

Description

The MAPK super family comprises c-Jun N-terminal kinases (JNKs), also known as stress activated protein kinases (SAPKs); extracellular signal-regulated kinases (ERKs); and p38 kinases, each of which forms a separate pathway. The kinase members the populate each pathway are sequentially activated by phosphorylation. Upon activation, p38 MAPK translocates into the nucleus (Figure 2) where it phosphorylates one or more nuclear substrates, effecting transcriptional changes and other cellular processes involved in cell growth, division, differentiation, inflammation, and death. p38 MAPK is activated by a variety of chemical stress inducers including hydrogen peroxide, heavy metals, anisomycin, sodium salicylate, LPS, and biological inducers such as certain growth factors and cytokines (e.g. Tumor Necrosis Factor-alpha, TNF-a).

The p38 MAPK Activation HCS Reagent Kit includes optimized and qualified high-content screening (HCS)-grade fluorescence reagents as well as a validated protocol to prepare and process samples for the quantification of p38 MAPK activation. Cells growing on standard high-density microplates are stimulated, and then fixed and immunofluorescently labeled to identify activated p38 MAPK in the cell nucleus (Figure 1). The antibody binds to dually phosphorylated p38 MAPK and has been validated for non-cross- reactivity to other MAPK/SAPK members as well as unphosphorylated p38 MAPK.

Prepared cells can be analyzed using standard fluorscence microscopy or using a Cellomics fully automated HCS Reader with the Cytoplasm to Nucleus Translocation Bioapplication, affording automated plate handling, focusing, image acquistion, analysis, and quantification and data storage.

Features

• A more biologically relevant measure of kinase activation relative to Western blots or in vitro kinase assays
• No cell lysis, purification, or filtration steps
• Immunofluorescence based assay provides sensitive results without radioactivity
• Kit antibody preferentially binds to dually phosphorylated (fully active) p38, with minimized cross-reactivity to other MAPKs
• Manual sample preparation/processing requires approximately 3 hours post-compound treatment

Related Poster Presentations

	Kay Opperman, Eric Hommema, Brian Webb, and Richik N. Ghosh (2008) Combining Multiplexed, High-Content Analysis with Antibody Arrays to Monitor and Correlate Intracellular and Extracellular Inflammatory Responses.
	Ulla Henriksen, Kay Opperman, Jacob Fog, Richik N. Ghosh, Frosty Loechel, Morten Praestegaard (2007) Profiling of Multiple Signal Pathway Activities by Multiplexing Antibody and GFP-based Translocation Assays.
	Ulla Henriksen, Kay Opperman, Jacob Fog, Richik N. Ghosh, Frosty Loechel, Morten Praestegaard (2007) Multiplexing of Antibody and GFP-based Translocation Assays: A Pathway to Better Profiling of Cellular Signalling.
	Chandrasekaran Vasudevan, Patrick Maunder, Jeffrey Haskins, Natalie Fursov Mei Cong and Zhong Zhong (2004) The Use of Division-Arrested Cells Improves the Robustness of High Content Screening Assays.
	Anthony Nichols, Francois Duval, Astrid Osen-Sand, Lilia Bernasconi-Castelli, Elisabeth Allet, Sandrine Pouly, Yves Sagot, Paul Lang and Alexander Scheer (2003) Use of Cellomics High Content Screening Platform to Increase Efficiency and Output for Small Molecule and Therapeutic Protein Discovery.

Kit Contents

• phospho-p38 MAPK primary antibody
• Alexa Fluor® 488 - conjugated secondary antibody
• Hoechst dye
• Block (100x)
• Wash buffer (10x)
• Permeabilization buffer (10x)
• Plate seals
  

Ordering Information

Screening size kits and components (bulk and custom) available with special pricing. Please call.

To order Cellomics HCS Reagent Kits, please call 800-874-3723 in the U.S.
or contact your local distributor of Thermo Scientific Products.

Choose "Order Online" in table to see pricing and order online (U.S. only).

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