simple, efficient, cell-based method to monitor proteins activated by phosphorylation

FACE  - Fast Activated proteins Cell-based ELISA Kits

• Metodo semplice, non radioattivo che puo' essere facilmente quantificato.

• Una rapida fissazione preserva le modiche della proteina attivata.

• Il saggio basato sulle cellule elimina la necessita' di fare gli estratti cellulari.

• Meno di tre ore effettive di lavoro.

• Pi quantitativo del Western Blot.

• High-throughput compatibile - 2 piastre e due anticorpi primari.

• Potente strumento di drug discovery.

simple, efficient analysis of cell function

Fast Activated Cell-based ELISA (FACE ) Kits provide a simple, efficient, cell-based method to monitor proteins activated by phosphorylation. FACE  ELISAs are 96-well, high-throughput assays that do not require cell extracts, electrophoresis or membrane blotting. Unlike Western blotting, FACE  enables modification-specific analysis inside the cell. This eliminates the need to perform cumbersome, low-throughput cell extractions and makes possible a 96-well format that is easily automated to screen large numbers of drug discovery compounds.

FACE  Kits are available for monitoring the cellular levels of activated p38, JNK, ERK1/2 and AKT, with many other kits on the way.

The FACE  Method

FACE  Kits are easy to use and require less than 3 hours of hands-on time. In FACE , cells are cultured in 96-well plates and stimulated to induce the pathway of interest. Following stimulation, the cells are fixed rapidly, which preserves activation-specific protein modifications. Each well is then incubated with a primary antibody specific for the activated protein of interest. Subsequent incubation with secondary HRP-conjugated antibody and developing solution provides a colorimetric readout that is quantitative and reproducible (Figure 1). The number of cells in each well can be normalized easily with the provided Crystal Violet solution. FACE  Kits also contain primary antibody specific for the native inactive protein, so you can monitor both native and activated protein levels in the same experiment.

FACE  eliminates cellular extractions, radioactive kinase assays, time-consuming Westerns and inefficient epitope interactions that occur on membranes. FACE  is a highly sensitive high-throughput assay designed for detecting activated proteins within mammalian cells.

Figure 1:
Flowchart of the FACE  process.