Drying of Polyacrylamide Gels

Drying Gels after staining with InstantBlue

InstantBlue is a ready-to-use, proprietary Coomassie®           
stain that is specially formulated for ultra-fast, sensitive
and safe detection of your proteins after electrophoretic
separation. Gels can be stained in minutes without the
need to wash, fix or destain. Long term storage of the
stained gels can be achieved by drying the gel post
staining with InstantBlue. An example of how this can be
achieved is discussed below.

• Staining the Gel

    Instant Blue is an ultra fast working stain. However although
    protein bands will be visible after a few minutes, the staining
    process is typically fully completed after 1h incubation. Depending
    on the type of gel you are using longer incubation may be
    necessary to complete the process. Further processing of the gel
    prior to completion of the staining process can result in protein
    destaining and reduced sensitivity. If this occurs simply restain
    the gel by incubating overnight in InstantBlue.

• Washing the Gel
    Prior to drying, unreacted stain must be removed from the gel. This is easily done by washing the  

    gel in water. By using hot water the speed of this process can be significantly increased.            

    Submersing a gel in 100 ml water, microwaving it for 1 minute at maximum power and incubating

    this solution for 1 hour typically removes most of the unreacted stain. Alternatively the gel can be

    kept overnight in ultrapure water at room temperature without loss of sensitivity.

• Conditioning the Gel
    To reduce the likelihood of cracks and uneven drying     
    submerse the gel in in a ‘gel drying solution’ (e.g. 4%
    glycerol, 20% ethanol in water) for maximum 5 minutes.
    Incubation of any Coomassie®-stained gel in an alcohol
    solution will eventually result in destaining of the bands so
    avoid incubation for longer than 5 minutes.

• Drying the Gel

    Insert the gel between two wetted cellophane membranes               
      and gently dry to further reduce the likelihood of cracking.
      Typically a gel can be dried within 48 hours.
 

Drying of Gels
Standard Protocol

    1. Remove the gel from the gel cassette.
        2. Submerse the gel in ~25 ml InstantBlue and incubate for 1 hour.
        3. Remove the gel from the stain, trim the edges and submerse in ~100 ml water.
            Microwave the submersed gel for 1 minute at maximum power and incubate for
            at least 1 hour in the hot water.
        4. Prepare ‘gel drying’ solution by mixing 4 ml glycerol with 76 ml ultrapure water
            and 20 ml ethanol (4% v/v glycerol, 20% v/v ethanol).
        5. Decant the water and add 100ml drying solution to the gel. Also submerse the
            cellophane membranes in the drying solution and incubate for 2 minutes.
        6. Install the drying frame on its holder and place one wetted membrane on the
            frame, followed by the gel and then the final membranes. Avoid air being
            trapped between the membranes and use additional drying solution if
            necessary.
        7. Close the frame and secure with clips. Stand the frame upright for 24 to 48
            hour to dry. Avoid drying too quickly as this increases the likelihood for
            cracking.
        8. Open the rack and remove the dried gel, trim the cellophane edges and stick in
            notebook for long term storage.