The STAT Activation assays are based on fixed end-point determinations and can be performed in multiple cell types. Following compound treatment, agonists are screened by immunofluorescence monitoring of the STAT isoform’s nuclear translocation. Whole cells are fixed, permeabilized, labeled with a STAT isoform specific primary antibody, and then stained with a secondary antibody conjugated with a fluorescent dye. Antagonist screens require the addition of a control inducer (e.g. interferon-a, IFN-g) to stimulate nuclear translocation of the STAT isoform prior to candidate drug treatment. Prepared cells can be analyzed using standard fluorescence microscopy or using Cellomics fully automated HCS Reader with the Cytoplasm to Nucleus Translocation BioApplication, affording automated plate handling, focusing, image acquisition analysis, quantification and data storage.

Features

• Kits detect STAT1, STAT2, or STAT3 activation specifically, sensitively, and reliably in whole cells without radioactivity
• Cost-effective, streamlined kits include all fluorescent reagents and protocols for optimized screening results
• No cell lysis, purification, or filtration steps
• Measurements made on endogenous STATs
• An earlier and more biologically relevant measure of transcription factor activation than gene reporter and electophoretic mobility shift assays

Kit Contents