Protein Work
Tips for Detecting Phosphoproteins
Protein phosphorylation is very important for protein post-translational modification that controls many cellular processes including metabolism, transcriptional and translation regulation, degradation of proteins, cellular signaling and communication, proliferation, differentiation, and cell survival [1].
Approximately 35% of human proteins are phosphorylated. Phosphoproteins are low in abundance, and, therefore, are, sometimes, challenging to detect and characterize by Western blot and other methods. Western blot is the most common method for detecting and quantifying phosphorylated proteins.
Quantitative and reliable protein 1D gel analysis demands a standardized and reproducible system. Proteins of samples separated by SDS- or IEF-PAGE are usually visualized using protein stains. Yet, protein stains are limited in their capabilities.
Due to the low detection sensitivity of Coomassie high amounts of sample are required. Both Coomassie and Silver stains provide a linear range of signals of only two orders of magnitude .
Fluorescent stains provide much better sensitivity and linear range, however, all stains suffer from limitations in terms of reproducibility and require additional time and labor for staining and destaining...