Improvements and simplifications in in-gel fluorescent detection of proteins using ruthenium II tris-(bathophenanthroline disulfonate): the poor man's fluorescent detection method

TL;DR

This paper presents improved, simplified protocols for in-gel fluorescent protein detection using ruthenium II tris-bathophenanthroline disulfonate, enhancing contrast and robustness while maintaining cost-effectiveness.

Contribution

The authors introduce modifications that simplify complex preparation and staining procedures, making the method more accessible and reliable for protein detection.

Findings

Higher contrast staining achieved

More robust complex preparation method developed

Maintains cost-effectiveness of the detection technique

Abstract

Fluorescent detection of proteins is a popular method of detection allying sensitivity, linearity and compatibility with mass spectrometry. Among the numerous methods described in the literature, staining with ruthenium II tris(bathophenanthroline disulfonate) is particularly cost-effective, but slightly cumbersome owing to difficulties in the preparation of the complex and complexity of staining protocols. We describe here the modifications on both aspects that allow to perform a higher contrast staining and offer a more robust method of complex preparation, thereby maximizing the advantages of the method.