Short-Run SDS-PAGE Protein Protocol

The objective of this method is to transfer a liquid-based solution to gel format. This permits easier sample manipulation, long-term storage and enables shipping at ambient temperature.

  1. Mix the sample with loading buffer and reducing buffer per the supplier's instructions.
  2. Heat at 90 C for 10 min.
  3. Allow to cool, spin and load the mixture to the well of a 16.5% Tris-Tricine gel. Do not use a gel with a stacking gel. We recommend loading a pre-stained molecular weight ladder standard in an adjacent well.
  4. Electrophorese at 100 V for about 10 min, or until the contents of the well have just entered the gel, as indicated by the loading buffer dye.
  5. Stop the electrophoresis, remove the cover plate from the gel and cut a single band from the lane containing the sample. The band should extend from the top of the lane (the bottom of the well) downward to a point indicated by the lowest band of the pre-stained ladder. When cutting the band use new gloves and a new scalpel blade (see our gel handling guide).
  6. Place the band into a micro-centrifuge tube pre-rinsed with MilliQ water.