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