User:Helen L. Slucher/Notebook/CHEM 571/2013/09/25

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Objective Run SDS-PAGE to observe amount of hemoglobin remaining after yesterday's digestion. Description Prepare the Gel and Assemble the Electrophoresis Cell Remove comb and tape from the gels Rinse the wells with running buffer Assemble the electrophoresis cell (note diagrams in manual) Fill the inner and outer buffer chambers with running buffer Prepare and Load Samples Heat your samples for 5 minutes at 100C (in the thermocycler) Load 20uL of protein ladder into column 1 of your gel Load 20uL of your samples into the appropriate lane of your gel Perform electrophoresis Run for 30 minutes at 200V (I need to make sure our power source can do this) Develop/Stain your gel Place gel in Fixative Solution (40% methanol, 10% acetic acid, 50% water) for 30 minutes Place gel in Stain Solution (0.025% (w/v) Coomassie Blue, 10% acetic acid, 90% water) for 1 hour Place gel in Destain Solution (10% acetic acid, 90% water) for 15 minutes Repeat this step with fresh destain solution 2 more times Data