Immunoblotting is frequently used for immunodetection of phosphoproteins, in some cases following immunoprecipitation of the cellular extract. The signals from each probe can then be compared and phospho-specific fractions visually assessed or quantified by densitometry. However, the time and effort required in these protocols has driven many researchers to seek out ELISA-based methods, which offer a faster, higher throughput alternative.
In this article, we present a study in which the phosphorylation of two proteins, STAT1 (Tyr701) and Mek1 (Ser217/221), is detected in response to various agonists. Data from the RayBio® Phosphorylation ELISA kits will be compared to traditional immunoblotting.
The detection of the phosphorylation of a specific protein is usually accomplished by immunological methods, in which the researcher can measure the fraction of phosphorylated target protein relative to total levels of expressed target protein in the cell. By comparing the phosphorylated fractions in cellular extracts under various experimental conditions, one ...
