Empirical Evidence of Cellular Histidine Phosphorylation by Immunoblotting Using pHis mAbs.

Empirical Evidence of Cellular Histidine Phosphorylation by Immunoblotting Using pHis mAbs. Methods Mol Biol. 2020;2077:181-191 Authors: Kalagiri R, Adam K, Hunter T Abstract Immunoblotting is a ubiquitous immunological technique that aids in detecting and quantifying proteins (including those of lower abundance) and their posttranslational modifications such as phosphorylation, acetylation, ubiquitylation, and sumoylation. The technique involves electrophoretically separating proteins on an SDS-PAGE gel, transferring them onto a PVDF (or nitrocellulose) membrane and probing with specific antibodies. Here we describe an immunoblotting technique for detecting cellular phosphohistidine, a labile posttranslational modification, by optimizing experimental conditions such that the labile phosphohistidine signal is conserved throughout the experiment. PMID: 31707659 [PubMed - in process]
Source: Mol Biol Cell - Category: Molecular Biology Authors: Tags: Methods Mol Biol Source Type: research