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Biology LibreTexts

14.4: Staining SDS-PAGE gels

  • Page ID
    17583
    1. Turn off the power supply.

    2. Remove the gel apparatus from the tank. Open the clamping frame and remove the gel cas- sette sandwich. Carefully, pry the two plates apart with a spatula. With the spatula, remove the lower right or left corner of the gel to serve as an orientation marker. Be sure to indicate in your lab notebook whether the notched corner corresponds to lane 1 or lane 10 of the gel.You may also remove the stacking gel with the spatula, if you desire.

    3. Place the gel in a small plastic tray and label the tray with your initials on a piece of tape. To do this, fill the tray about halfway with deionized water. Gently free the gel from the glass plate, allowing it to slide into the water. The gel should move freely in the water. Place the gel and tray on a rocking platform. Rock the gel for ~2 minutes.

    4. Drain the water from the gel and add enough Simply Blue to cover the gel, while allowing the gel to move freely when the tray is rocked. Cover the gel container with saran‐wrap and rock overnight. Make sure that the gel does not stick to the bottom of the tray.

    5. In the morning, drain the Simply Blue stain into an appropriately labeled waste container in the hood of the lab room.

    6. Destain the gel by filling the container about half full with deionized water. Shake the gel in the water for ~2 minutes. Pour off the water and add new deionized water. Repeat, if necessary, until protein bands become visible.

    7. When individual bands are detectable, record your data. You may photograph the gel with your cell phone camera against a white background. Alternatively, place the gel in a clear plastic page protector and scan the gel.

    8. After recording the data, dispose of the gel in the Biohazard waste container.