Cell Lysates Protocol

Lysates are supplied at a protein concentration of 1 mg/mL in non-reducing 1X SDS-PAGE loading buffer.

Store at -80 °C.

If the entire unit of cell lysate will not be used at one time, thaw and divide into aliquots of a convenient size. Store aliquots at -80 °C. To prepare the lysate for SDS-PAGE, first thaw the appropriate volume of lysate. If reducing conditions are desired, add beta-mercaptoethanol to 4% and mix well. Boil the lysate for 5 minutes followed by centrifugation at high speed for 3 minutes.

Load 35 µL (35 µg) of lysate per well of the SDS-polyacrylamide gel. Following electrophoresis, transfer the resolved lysate proteins to nitrocellulose.