Fluorescent Silver Staining: A Technique to Visualize Total Protein in Polyacrylamide Gels

Published: April 30, 2023

Abstract

Source: Wong, A. Y. H. et al. Fluorescent Silver Staining of Proteins in Polyacrylamide Gels. J. Vis. Exp. (2019)

In this video, we demonstrate a silver staining method of polyacrylamide gels for the visualization of total protein by using fluorogenic probes.

Protocol

1. Preparation of the Gel Perform SDS-PAGE with 4% – 12% Bis-Tris protein gels (1 mm, 15-well) using a mini gel tank filled with 2-(N-morpholino)ethanesulfonic acid (MES) buffer. Dilute the samples with a mixture of distilled water, lithium dodecyl sulfate (LDS) buffer, and a sample-reducing agent. Load the first lane with double the amount of stock (10 µL), followed by the normal stock amount (5 µL) and a series of twofold dilutions of the stock thereafter (13 dilutions…

Divulgations

The authors have nothing to disclose.

Materials

LDS Sample Buffer (4X) Thermo Fisher Scientific NP0007 Reagent
4-12% Bis-Tris Protein Gels, 1.0 mm, 15-well Thermo Fisher Scientific NP0323BOX Precast gel
Sample Reducing Agent (10X) Thermo Fisher Scientific NP0004 Reagent
MES SDS Running Buffer (20X) Thermo Fisher Scientific NP0002 Reagent
Mini Gel Tank Thermo Fisher Scientific A25977 Equipment
300W Power Supply (230 VAC) Thermo Fisher Scientific PS0301 Equipment
Unstained Protein Ladder Thermo Fisher Scientific 26614 Sample
Silver nitrate Sigma-Aldrich 31630-25G-R Reagent
Ethanol Bragg and co. 42520J Reagent
Acetic acid J.T. Baker 103201A Reagent
Milli-Q Synthesis A10 Merk Provides 18.2 MΩ.cm water
Gel documentation system (c600 model) Azure biosystems Equipment

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Citer Cet Article
Fluorescent Silver Staining: A Technique to Visualize Total Protein in Polyacrylamide Gels. J. Vis. Exp. (Pending Publication), e21043, doi: (2023).

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