Brilliant Blue G – Colloidal Concentrate Protocol

Product No. B2025

Product Description

Brilliant Blue G-Colloidal Concentrate has been designed for post-electrophoresis staining of proteins in IEF, PAGE, and SDS-PAGE gels. Fixing the proteins prior to staining is recommended for maximum sensitivity.

Reagents and Equipment Required but Not Provided

• Fixing Solution (F7264)
• Acetic Acid, Glacial (A6283)
• Methanol (M3641)
• Ammonium Sulfate (A4915)

Precautions and Disclaimer

This product is for R&D use only, not for drug, household, or other uses. Please consult the Safety Data Sheet for information regarding hazards and safe handling practices. 

Procedure

1. After electrophoresis, fix the proteins for 30 minutes in Fixing Solution or one hour in a solution of 7% glacial acetic acid in 40% (v/v) methanol.
2. Add 800 mL of deionized water to the bottle labeled Brilliant Blue G-Colloidal Concentrate. Replace the cap and tighten. Mix by inversion. This 1X working solution should not be filtered. Store at 2-8 °C once diluted.
3. Immediately before staining, combine 4 parts of the 1X working solution (from step 2) and 1 part methanol. Mix well by vortexing for 30 seconds.
   NOTE: If the working solution was not freshly prepared, mix by inversion prior to combining with methanol. The staining suspension containing methanol is stable for 4 hours. Add methanol only to the amount of working solution that will be used at once.
4. Place the gel in staining suspension (from step 3) for 1-2 hours.
5. Destain with 10% acetic acid in 25% (v/v) methanol for 60 seconds with shaking. For gels <1.5 mm thickness, reduce this destaining time to 10-30 seconds.
6. Rinse the gel with 25% methanol, discard, and then destain in 25% methanol for up to 24 hours. If any precipitated dye remains on the surface of the gel, gently wipe with a clean cotton ball or a lab wipe soaked in 25% methanol.
7. Scan gel at 600 nm.
8. Gels may be stored for several weeks in 25% (v/v) ammonium sulfate at room temperature.

References

1. Neuhoff V, Arold N, Taube D, Ehrhardt W. 1988. Improved staining of proteins in polyacrylamide gels including isoelectric focusing gels with clear background at nanogram sensitivity using Coomassie Brilliant Blue G-250 and R-250. Electrophoresis. 9(6):255-262. https://doi.org/10.1002/elps.1150090603