Protein Expression Analysis Question - Protein Expression Analysis (Jul/26/2005 )

Hi,

I am fairly new to the world of protein purification and I have been given 2 clones which I am to test for protein expression. What is the best way to test for protein expression after induction with 1mM IPTG.

I should run SDS page to test for expression. What do I do to these cells after induction to prepare them for running on the gel.

Thanks.

JSE

For analysis of protein expression after IPTG induction you need to sonicate the cells and run the clarified ( by centrifugation) sample along with uniduced sample as control. However, you may not see an additional band conspicuously if the expression is not very high. Distinct bands appear when expressed in pET/BL21 system.

Other possibilities are:

1. Biological activity assay , if the overepxressed proein is an enzyme.
2. Westernblotting followed by staining with specific antibodies.