fluorescent-silver-staining-of-proteins-in-polyacrylamide-gels

Fluorescent Silver Staining of Proteins in Polyacrylamide Gels

Silver staining is a colorimetric technique widely used to visualize protein bands in polyacrylamide gels following sodium dodecyl sulfate-polyacrylamide ...

Karolinska Institutet

Silver staining, which exploits the special bioaffinity and the chromogenic reduction of silver ions, is an indispensable visualization method in biology. It is a most popular method for in-gel protein detection. However, it is limited by run-to-run variability, background staining, inability for protein quantification, and limited compatibility with mass spectroscopic (MS) analysis; limitations that are largely attributed to the tricky chromogenic visualization. Herein, we reported a novel water-soluble fluorogenic Ag probe, the sensing mechanism of which is based on an aggregation-induced emission (AIE) process driven by tetrazolate-Ag interactions. The fluorogenic sensing can substitute the chromogenic reaction, leading to a new fluorescence silver staining method. This new staining method offers sensitive detection of total proteins in polyacrylamide gels with a broad linear dynamic range and robust operations that rival the silver nitrate stain and the best fluorescent stains.

Fluorogenic Ag -Tetrazolate Aggregation Enables Efficient Fluorescent Biological Silver Staining.

Protein is one of the four most important biomacromolecules in living systems. The detection, quantification, localization, and characterization of proteins is essential for an understanding of biological fundamentals, as well as for the diagnostics and treatment of protein-related diseases. By using intrinsic and extrinsic fluorescence, different techniques have been established to study proteins, many of which are now being routinely used in research laboratories and clinics. This review summarizes the applications of aggregation-induced emission (AIE) fluorescence in protein science. In contrast to traditional fluorescent dyes, the activation of AIE dyes is mainly attributed to the restriction of intramolecular motions. This unique turn-on mechanism of AIE dyes allows researchers to develop novel fluorogenic strategies for sensitive, selective, and reliable analysis of proteins. This review focuses on introducing AIE strategies for 1) detection, localization, and quantification of proteins; 2) probing polymer conformational transitions of proteins; 3) characterization of protein-ligand interactions; and 4) evaluation of enzyme activities. Perspectives and challenges with respect to this emerging field of protein characterization are offered.

Alex Y. H. Wong

Ming Wai Lau Centre for Reparative Medicine

Karolinska Institutet

Fluorescent Silver Staining of Proteins in Polyacrylamide Gels

Alex Y. H. Wong

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Karolinska Institutet

Fluorescent Silver Staining of Proteins in Polyacrylamide Gels

Ming Wai Lau Centre for Reparative Medicine