ATF-2 Activation Kit

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 ATF-2 Activation Kit
 Product# Price  Product Name  Pack Size  
         
 K0100101      -    ATF-2 (green) Activation  5 x 96    SELECT   
 R0105141      -    ATF-2 (green) Activation  50 x 96    SELECT   
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  Details All Product Numbers
 

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Figure 2. Diagrammatic view of the principle of the ATF-2 Activation Reagent Kit.  ATF-2 (green), normally present in the cytoplasm, translocates to the nucleus.

Figure 1. Images of immunofluorescently stained NIH 3T3 cells before (left) and after (right) activation by ATF-2 by anisomycin. Cells were stimulated with 200 ng/mL anisomycin for 30 min and then prepared using the kit protocol. Nuclei appear blue (Hoechst) and ATF-2 appears green. Images were aquired using the ArrayScan? HCS Reader.

The ATF-2 Activation Kit, in combination with the Thermo Scientific ArrayScan? HCS Reader and the Cytoplasm to Nucleus Translocation or Molecular Translocation BioApplication software, enable automated plate handling, focusing, cell image acquisition, analysis and quantification of ATF-2 activation.

The ATF-2 transcription factor is activated in response to inflammatory cytokines and cellular stress, including genotoxicity and ischemia/reperfusion. ATF-2 is activated through phosphorylation of threonines 69 and 71 by members of the SAPK family. Once activated, ATF-2 forms complexes with Jun family or other ATF family members. These complexes bind to the cAMP response element (CRE) present in the promoters of many genes, stimulating gene transcription. Activation of ATF-2 can be quantified by its translocation to the nucleus. The immunofluorescent assay uses a validated phospho-ATF-2 specific antibody that is non-reactive to other cAMP response element-binding protein (CREB) or ATF family members.

Cell-based assay provides an earlier and more definitive measure of ATF-2 activation relative to electophoretic mobility shift or gene reporter assays
Cells prepared with the HCS Reagent Kits may be analyzed directly via standard fluorescence microscopy; or any HCS Imaging System.
Prepared cells are ready for analysis in less than 3 hours post-test compound incubation; kit protocol requires no cell lysis, purification or filtration steps
Immunofluorescence-based screening assay provides sensitive quantitative and qualitative results without radioactivity
Validated protocol included with all necessary stains and components
The Cellomics ATF-2 Activation Kit enables quantitation of ATF-2 activation by directly measuring its translocation from the cytoplasm to the nucleus. The assay is performed on live cells growing on standard high density microplates. The kit is supplied with a phospho-ATF-2 specific antibody and a DyLight 488-conjugated Secondary Antibody. The nuclear region is identified by Hoechst Dye, which is also included.

 
ATF-2 Activation Kit Features

   Product Detail 

The Activating Transcription Factor-2 (ATF-2) Activation HCS Reagent Kit provides high-content screening (HCS)-quality fluorescence reagents and a validated, optimized protocol for measuring and visualizing the activation of the ATF-2 transcription factor.