Silver Staining of Sulfated Glycosaminoglycans: A Staining Method for Visualization of Sulfated Glycosaminoglycans in Polyacrylamide Gels

Abstract

Source: LaRiviere, W. B., et al. Detection of Glycosaminoglycans by Polyacrylamide Gel Electrophoresis and Silver Staining. J. Vis. Exp. (2021)

In this video, we demonstrate a procedure for combined Alcian blue and silver staining of sulfated glycosaminoglycan oligosaccharide fragments. This combined staining technique significantly amplifies the bands of glycosaminoglycan polymers of different lengths in polyacrylamide gels.

Protocol

1. Silver staining protocol Prepare all solutions necessary for silver staining in advance (Table 1). NOTE: Do not directly touch the PAGE gel until it has been stained, developed, and placed in stop solution. Instead, manipulate the gel using clean plastic or glass tools. Directly handling the gel will result in finger-print distortions and other visible artifacts on the gel after staining. Once th…

Materials

Alcian Blue 8GX (solid)	ThermoFisher Scientific	AC400460100
Ammonium hydroxide (liquid)	ThermoFisher Scientific	A669S-500	Certified ACS
Barnstead GenPure Pro Water Purification System	ThermoFisher Scientific	10-451-217PKG	Any water deionizing/ purification system is an acceptable substitute
Glacial acetic acid (liquid)	ThermoFisher Scientific	A35-500	Certified ACS
Methanol (liquid)	ThermoFisher Scientific	A412-500	Certified ACS
Silver nitrate (solid)	ThermoFisher Scientific	S181-25	Certified ACS
Sodium hydroxide (solid)	ThermoFisher Scientific	S392-212
Vacuum filter unit, single use, 0.22uM pore PES, 500mL volume	ThermoFisher Scientific	569-0020	Alternative volumes and filter materials acceptable
Molecular Imager Gel Doc XR System	Bio-Rad	170-8170	Any comparable gel imaging system is an acceptable substitute