The NFAT Activation HCS Reagent Kit provides for high-content screening (HCS)-quality fluorescence reagents along with a validated, optimzied protocol for measuring and visualizing the activation of the NFAT transcription factor. The assay is performed on live cells growing on standard high-density microplates, and is based on immunofluorescence, using and NFAT-specific antibody and a fluorophore-conjugated secondary antibody. Hoechst Dye, also included in the kit, identifies the nuclear region of each cell.

The NFAT Activation HCS Reagent Kit has been optimized for a fixed end-point assay. Inhibitors of NFAT translocation are screened by stimulating cells with a control compound such as thapsigargin following the exposure of live cells to the test compounds. Replacing the control compound with test compounds identifies agonists of NFAT translocation, which is directly quantified as the difference in cytoplasmic to nuclear intensity of the labeled transcription factor (Figures 1 & 2). Because NFAT must be in the nucleus to induce gene expression, its translocation is a definitive measure of its activation, but marks an earlier event than reporter gene expression. Further, NFAT translocation represents a functional, more biologically relevant assay in comparison to gel mobility shift or gene reporter assays.

Kit Contents

• NFAT 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).

See all Cellomics HCS Reagent Kits