Research detection kit HTRF

total proteinscell

This HTRF kit detects cellular STAT6 and can be used as a normalization assay with our phospho-STAT6 kit for an optimal readout of JAK/STAT signaling. OverView The Total STAT6 cellular assay kit is designed to monitor the expression level of STAT6, whether phosphorylated or unphosphorylated. It is compatible with our Phospho-STAT6 kit, and enables the analysis of phosphorylated and total proteins from a single sample for better readouts of the JAK/STAT signaling pathway. Specifications Other Specifications Unit Size 500 Assay Points How it works Total STAT6 assay principle The Total STAT6 assay quantifies the expression level of STAT6 in a cell lysate. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer. The Total-STAT6 assay uses two labeled antibodies: one coupled to a donor fluorophore, the other to an acceptor. Both antibodies are highly specific for a distinct epitope on the protein. In presence of STAT6 in a cell extract, the addition of these conjugates brings the donor fluorophore into close proximity with the acceptor and thereby generates a FRET signal. Its intensity is directly proportional to the concentration of the protein present in the sample, and provides a means of assessing the proteins expression under a no-wash assay format. Total STAT6 2-plate assay protocol The 2-plate protocol involves culturing cells in a 96-well plate before lysis, then transferring lysates to a 384-well low volume detection plate before the addition of Total-STAT6 HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.