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

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…

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