Introduction:

Coomassie Brilliant Blue Staining Solution (destaining-free) is a non-toxic and non-irritating odor coomassie G-250 dye-based reagent for instant staining protein on polyacrylamide gels. It offers the following advantages over conventional colorimetric stains:

• More convenient: No need to wash, fix, heat or destain.
• High sensitivity: Detect protein bands as low as 10ng.
• Ultra-fast staining: Protein bands can be visualized in 5 minutes.
• Clear background: High signal-to-noise ratio.
• Good compatibility: Suitable for denaturing and non-denaturing gels and mass spectrometry.
• Ingredient safety: Does not contain methanol, acetic acid and other toxic and irritating substances. No need to use a fume hood.  

Storage: Store at room temperature, valid for 1 year.

Protocol (For 0.75 mm thickness, 8×10 cm gel, for reference only):

1. It is normal to find a little precipitation in the bottle before use, which will not affect the dyeing effect. Please mix thoroughly before use.
2. Remove the PAGE gel after electrophoresis and put it into a plastic container, add an appropriate volume of staining solution to completely cover the PAGE gel. The volume of staining solution depends on the size of the container, usually about 30-50mL.
3. Place the gel container on a shaker for gently shaking. Protein bands will begin to appear after 2-3 minutes and deepen over time. Staining time can be from 30 minutes to overnight. Background color will not substantially increase with staining time.

Note: Since the dye solution is acidic, do not touch the dye solution with bare hands. The dye solution can be reused 3 times. As the number of uses increases, the staining speed decreases slightly, but the sensitivity does not decrease significantly.

4. After staining, store the gel in water for subsequent photograph. Due to the different isoelectric points of proteins, the instantaneous adhesion ability of dyes is different. It is recommended to place the gel in water after staining for 2 hours or overnight.

Features:

The stain solution only stains the protein but not the gel background, and no destaining is required. The staining speed is fast and the next experimental steps can be determined according to the results of protein staining bands within a few minutes after staining. Simple to operate, easy to store, no special equipment is required, and the gel can be stored in pure water for several months.

Scope of application:

This product is suitable for the staining hand-made or precast SDS-PAGE gels and Native gels, and can also be used to stain residual proteins on PAGE gels after western transfer.

Notes:

1. The various parameters in the protocol are only for the gel with an area of 8 × 10 cm and a thickness of 0.75 mm. If you use a larger area or thicker gel, please add more staining solution and prolong the staining time.
2. If the gel needs to be completely free of background, it can be washed with distilled water until the background is clean, no need to use destaining solution.
3. Like the traditional Coomassie brilliant blue staining solution, this stain solution binds positively charged groups on proteins. The quantitative comparison between different proteins needs to consider the number of positively charged amino acids. Some proteins stain easily, while others stain less. This is related to the isoelectric point of the protein and the amount of lysine and arginine contained in it.
4. The stain solution is acidic and slightly corrosive. For your safety and health, please wear a lab coat and disposable gloves.
5. For research use only. Not for use in diagnostic procedures.

1. Zengwen Huang, Juan Zhang, Yaling Gu, Zhengyun Cai, Dawei Wei, Xiaofang Feng, and Chaoyun Yang. Analysis of the molecular mechanism of inosine monophosphate deposition in Jingyuan chicken muscles using a proteomic approach. Poultry Science 101, 101741, April 2022. （doi.org/10.1016/j.psj.2022.101741） (IF=4.2)