Quick Coomassie Staining System - Code J558

Directions for use

The AMRESCO Quick Coomassie Staining System is intended for staining of proteins after SDS-PAGE. The system is designed to yield completely stained and destained protein bands in 60 minutes or less. After reconstitution, the Quick Coomassie Staining System contains sufficient reagents to stain 40 standard mini-gels.

a. J558-40Gels have

• 400 ml Coomassie Reagent 1

Concentrate (Code J534)

• 400 ml Coomassie Reagent 2

Concentrate (Code J539)

OR

b. J558-Q-4Gels have

• 40 ml Coomassie Reagent 1

Concentrate (Code J534)

• 40 ml Coomassie Reagent 2

Concentrate (Code J539)

• Methanol (Code 0323)

• Acetic Acid, Glacial

• Destain Solution 1

  (Prepare fresh each time)

  200 ml Deionized Water

  250 ml Methanol (Code 0323)

  50 ml Acetic Acid

  Yields 500 ml

• Destain Solution 2

  (Prepare fresh each time)

  440 ml Deionized Water

  25 ml Methanol

  35 ml Acetic Acid

  Yields 500 ml

• Agitate both bottles of Quick Coomassie Reagent Concentrate briefly to ensure no settling of reagents has occurred during shipping.

• NOTE: When using J558-40Gels, please follow Step 2.a. When using J558-Q-4Gels, please follow Step 2.b.

  • To each bottle of Reagent Concentrate, add 500 ml Methanol and 100 ml Acetic Acid. Cap each bottle and invert briefly to mix. This reconstituted material should be stored at room temperature (18- 26°C) and used within 3 months.

  or

  • To each bottle of Reagent Concentrate, add 50 ml Methanol and 10 ml Acetic Acid. Cap each bottle and invert briefly to mix. This reconstituted material should be stored at room temperature (18- 26°C) and used within 3 months.

• After gel electrophoresis is complete, disassemble the plates and transfer the gel to a clean, plastic or glass container. To minimize waste, it is best to choose a container only slightly larger than the gel to be stained. See Notes and Tips for Success Section.
• Determine the volume of stain required to completely cover the gel. (In most cases, a standard mini-gel can be stained with 50 ml of Staining Solution.) For each 50 ml of stain required, add 25 ml of reconstituted Quick Coomassie Reagent 1 and 25 ml of reconstituted Quick Coomassie Reagent 2. [NOTE: Do not mix Quick Coomassie Reagent 1 and Quick Coomassie Reagent 2 together until ready to use].
• Allow the gel to stain, with shaking, for 10 minutes. [NOTE: Longer staining periods will not increase stain sensitivity and will result in longer destain times].

• Decant and discard Staining Solution and add 3 volumes of Destain Solution 1. Incubate, with gentle shaking, 5 minutes

• Replace Destain Solution 1 with fresh Destain Solution 1 and incubate, with shaking, an additional 5 minutes. Discard Destain Solution 1.
• Add 3 volumes of Destain Solution 2, and destain gel until desired background is achieved. In most cases, bands are fully visible after 15 minutes and background will be completely clear after 30-60 minutes.

• Preparing fresh Destain Solution 1 and Destain Solution 2 each time is essential to minimizing your destaining times. Older, stored destaining solutions may result in longer total destaining times.

• Pay particular attention to the stain time in Step 5. Staining times longer than 10 minutes may result in over-stained gels that will require longer destaining periods to achieve clear background.

• Regarding the size of the staining container: To minimize waste, choose a container large enough to allow movement of the staining and destaining solutions around the gel, but small enough to minimize the amount of solution required to cover the gel. A standard, 10X10 cm mini-gel is best stained on one half of an empty 1000 µL pipette tip ("Blue Tip") box.

• Do not attempt to re-use the staining solution. Attempting to do so may result in greatly decreased stain sensitivity.

• Some users find they can decrease destaining times even further by microwave-treating the gel in Destain Solution I. After removing the Staining Solution, simply add Destain Solution 1 and microwave on full power for 15 seconds, prior to the first 5-minute incubation step.