YOUR CART
The Forkhead transcription factor FKHR (FOXO1) regulates expression of gluconeogenic factors, and plays a key role in the insulin regulation of glucose levels. Besides its role in glucose homeostasis, FKHR has been shown to be involved in pathological processes such as type 2 diabetes, cancer and organismal aging. The transcriptional output of FKHR is controlled by a two-tiered mechanism of phosphorylation and acetylation, and alterations in this balance can result in profound effects.
The FACE™ Method
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 or chemiluminescent 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.
