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

Published: April 30, 2023

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…

Açıklamalar

The authors have nothing to disclose.

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

Etiketler

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Bu Makaleden Alıntı Yapın
Silver Staining of Sulfated Glycosaminoglycans: A Staining Method for Visualization of Sulfated Glycosaminoglycans in Polyacrylamide Gels. J. Vis. Exp. (Pending Publication), e21049, doi: (2023).

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