Method Article

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

July 8th, 2025

In This Article

Abstract

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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

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1. Silver staining protocol

  1. 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.
  2. Once the run is completed, disassemble cassette and extract gel in a clean, medium-large container filled with deionized, filtered water.
    NOTE: To avoid directly hand....

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Disclosures

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No conflicts of interest declared.

Materials

List of materials used in this article
NameCompanyCatalog NumberComments
Alcian Blue 8GX (solid) ThermoFisher Scientific AC400460100
Ammonium hydroxide (liquid) ThermoFisher Scientific A669S-500 Certified ACS
Barnstead GenPure Pro Water Purification SystemThermoFisher 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 volumeThermoFisher Scientific 569-0020 Alternative volumes and filter materials acceptable
Molecular Imager Gel Doc XR SystemBio-Rad170-8170 Any comparable gel imaging system is an acceptable substitute

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Tags

Alcian Blue StainingPolyacrylamide Gel ElectrophoresisGlycosaminoglycan VisualizationAcidic Alcian BlueAlkaline Silver NitrateDeveloping Solution FormaldehydeStop Solution pHGel Washing Procedure

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