Non-Reducing SDS PAGE: A Method to Analyze Disulfide-Linked Multimeric Protein Complexes
Transcript
Prepare 1 liter of 1X Tris-glycine running buffer by mixing 25 millimolar Tris, 192 millimolar glycine, and 0.1% weight per volume SDS. Then, set up the SDS-PAGE running apparatus.
Open the 16% pre-cast TGX SDS-PAGE package per the manufacturer's protocol, and remove the cassette. Remove the comb that is lining the wells, and the tape from the bottom of the cassette, and place the gel into the running apparatus. Fill the chamber with the 1X running buffer, until the wells are submerged in liquid.
Using a plastic pipette, rinse out the wells with the running buffer. Load the samples onto the gel along with 10 microliters of the pre-stained standard marker. Finally, run the gel at 200 volts until the dye front is approximately 1 centimeter from the bottom of the gel.
