SDS-PAGE

SDS-PAGE Acronym

Sodium Dodecyl Sulfate Poly-Acrylamide Gel Electrophoresis

How proteins move and separate in SDS-PAGE

1. Proteins are negatively charged and move towards the anode (+)

2. That are separated based on size

SDS

1. A detergent that denatures proteins

   1. Breaks non-covalent bonds

2. Coats proteins to give them a net negative charge

3. Know SDS is present in a solution if there are bubbles

Reducing agents in SDS-PAGE

BME and DTT

BME and DTT

1. Reducing agents that help denature proteins

   1. Breaks covalent disulfide bonds

Heat in SDS-PAGE

1. Helps denature proteins

2. 95-100°C for 3-5 (few) min.