Detection and quantification of C-terminally tagged proteins by in-gel fluorescence.

ABSTRACT: The analysis of recombinant proteins in complex solutions is often accomplished with tag-specific antibodies in western blots. Recently, I introduced an antibody-free alternative wherein tagged proteins are visualized directly within polyacrylamide gels. For this, I used the protein ligase Connectase to selectively attach fluorophores to target proteins possessing an N-terminal recognition sequence. In this study, I extend this methodology to encompass the detection and quantification of C-terminally tagged proteins. Similar to the N-terminal labeling method, this adapted procedure offers increased speed, heightened sensitivity, and an improved signal-to-noise ratio when compared to western blots. It also eliminates the need for sample-specific optimization, enables more consistent and pre

SUBMITTER: Fuchs ACD 

PROVIDER: S-EPMC11231263 | biostudies-literature | 2024 Jul

REPOSITORIES: biostudies-literature