To run an SDS-PAGE to determine whether or not MBP-intein fusion proteins are being expressed cells comparable to the cells frozen on 09/28/11.
- Make solutions for a 12% discontinuous gel using the BioRad Mini-PROTEAN instructions.
- Set resolving gel solution in gel holder with methanol on top to make the top of the gel flat.
- Allow gel to sit for 20 min.
- Remove methanol.
- Add stacking gel solution.
- Load gels into SDS-PAGE.
- Load ladder and protein solutions into gel.
- Run SDS-PAGE.
- Stain gels with Coomassie Brilliant Blue R250 solution.
- Dissolve 0.2555 g Coomassie Brilliant Blue R250 in 90 mL of methanol:water (1:1 v/v) and 10 mL of glacial acetic acid.
- Filter solution through a Whatman No. 1 filter.
- Place on shaker overnight.
Protein visibly traveled on gel after SDS-PAGE.
Protein solutions used were from group 2: