Cellomics HCS Reagent Kit for Protein Kinase C Alpha (PKCa).

Description

The Protein Kinase C (PKC) family is a group of related signal transduction proteins, and their activation is one of the earliest events leading to a variety of cellular responses including cytokine secretion, differentiation, proliferation, muscle contraction and modulation of membrane excitability. PKCa is believed to play an important role in the development and growth of cancer cells. Inhibition of PKCa production is the basis for therapeutic candidates currently in clinical trials for the treatment of non-small cell lung cancer. PKCa may also underlie the cellular mechanism leading to inflammatory lung vascular injury and adult respiratory distress syndrome.

The PKCa Activation HCS Reagent Kit comprises optimized and qualified high-content screening (HCS)-grade fluorescence reagents and an automation-compatible protocol for the quantification of PKCa translocation. Cells growing on standand high-density microplates are stimulated, fixed and immunofluorescently labeled to identify PKCa (Figure 1). The kit also includes a cell membrane marker so co-localization of PKCa at the cell membrane may be quantified. Based on indirect immunofluorescence detection of the translocation of PKCa between cytoplasm and plasma membrane, this kit may be used to screen for inhibitors, activators, or modulators of PKCa.

Fully automated imaging and analysis of the prepared samples may be performed using a Cellomics fully automated ArrayScan HCS Reader with the Cyto-Cell Membrane Translocation BioApplication, which affords automated plate handling, focusing, image acquistion, quantification, and data storage.

Features

• Time-saving and convenient kit provides HCS-qualified antibodies and fluorescent reagents that identify the cell membrane, nucleus, as well as PKCa in cells
• More physiologically relevant information provided through measurement of endogenous PKCa
• Convenient automation-compatible protocol requires no cell lysis, protein purification, generation of stable transfected cell lines, or radioactivity
• Cells prepared with the HCS Reagent Kits may be analyzed directly via standard fluorescence microscopy; or HCS imaging systems

Related Poster Presentations

	Richard DeBiasio, Richik N. Ghosh, Alexander L. Friedman and Jeffrey R. Haskins (2004) Optical Sectioning With the Apotome Attachment on the ArrayScan HCS Reader Enhances High Content Screening Assays.
	Yih-Tai Chen, Kenneth A. Guiliano, and Jeffrey R. Haskins (2003) High-Content Screening of Gene Knock-downs Induced with siRNA Transfection.
	Jeffrey R. Haskins, Chandrasekaran Vasudevan, Patrick Maunder and Yih-Tai Chen (2003) Positional Biosensors Enable Large-Scale Investigation of Apoptotic Mechanisms of Cell Death.
	Cellomics, Inc. (2003) Unlocking the Power of Optical Sectioning with the ArrayScan V HCS Reader.
	Yih-Tai Chen, Kenneth A. Giuliano, and Jeffrey R. Haskins (2003) Validation of siRNA Gene Knockdown Through Automated Phenotypic Analysis of Cellular Function.

Kit Components

• PKCa monoclonal antibody
• Secondary antibody Alexa Fluor® 555 conjugate
• Membrane marker-Alexa Fluor® 488 conjugate
• Hoechst dye
• Block (100x)
• Wash buffer-M (10x)
• Permeabilization buffer (10x)
• Detailed protocol and assay data
• 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).

See all Cellomics HCS Reagent Kits