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  • Using NEB Protein Markers & Ladders

    Thawing

    • Markers and Ladders should be thawed on ice

    Loading a sample

    • Vortex gently to ensure the solution is homogenous
    • Aliquot the following amount into a separate microcentrifuge tube:
      Marker/Ladder mini gel full-size gel
      Protein Marker, Broad Range 7 µl 15 µl
      Protein Ladder 5 µl 10 µl
      Prestained Protein Marker 6 µl 12 µl (blotting)
        15 µl 30 µl (visualizing)
      ColorPlus Prestained Protein Marker 6 µl 12 µl (blotting)
        15 µl 30 µl (visualizing)
    • Heat sample at 95–100°C for 3–5 minutes
    • After a quick microcentrifuge spin, load directly onto a gel
    • To ensure uniform mobility, load an equal volume of 1X SDS loading buffer into unused wells

    Storage

    • Recommended storage for all protein markers and ladders is -20°C

    Analysis

    • The Protein Ladder (NEB #P7703) and the Protein Marker, Broad Range (NEB #P7702) give their true molecular weight values when run on a gel. For accurate size determination, the Protein Ladder is recommended.
    • “Apparent” molecular weights are assigned to the Prestained Protein Markers (NEB #P7708, #P7709). These markers are not recommended for size determination. When a protein is covalently bound to a charge-carrying dye molecule, the protein’s overall charge can be affected and alter its mobility within the gel. In addition, prestained protein markers will resolve differently on varying gel types.

    (From NEB expressions Spring 2008, vol 3.1)