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
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).