Suggested Protocol for Loading the Unstained Protein Ladder, Broad Range (P7703)

1. Gently mix the Unstained Protein Ladder, Broad Range (NEB #P7703 ) by gently vortexing or pipetting up and down several times.
2. Incubate at 95°C-100ºC for 5 minutes.
3. Centrifuge quickly.
4. Aliquot in to single use aliquots and store at -20C. Do not boil the aliquots again.
5. Load 5 μl of P7703 in a gel lane.
6. To ensure uniform mobility, load an equal volume of 3X Blue SDS Loading Buffer (NEB #B7703) into any unused wells.
7. Run gel using constant voltage, according to manufacturer’s specifications.
8. Run until dye front reaches bottom of gel, but does not run off.
9. Stain and de-stain gel with protein stain of choice, following manufacturer’s instructions (i.e. Coomassie Brilliant Blue, Simply Blue^TM Safe Stain, or GelCode^TM Blue Safe Stain)

Note: Do not boil the aliquots a second time. The recommended load for (NEB #P7703) is 5 μl. Loading less than this amount may result in less intense bands.