Christopher Gaughan

3 Questions 5 Answers 0 Followers

Questions related from Christopher Gaughan

Has anyone ever found that PMSF can affect enzyme activity of recombinantly expressed proteins?

I am expressing 2 enzymes in BL21, one is the WT form and the other a variant. I expressed the WT form without using any PMSF in the lysis buffer and the activity is fine. However, when I express...

14 August 2013 5,744 12 View

Must I add RNAse and DNAse to my lysis buffers for native purification in E coli?

I will be expressing three soluble proteins in E. coli BL-21-T1 cells (Sigma competent cells). The plasmid has a T5 promoter and is induced by IPTG. Many investigators site that the lysis step is...

10 July 2013 8,481 9 View

Has anyone used the Lightening Link Product (Innova Biosciences) to link PerCp to proteins other than antibodies? I am looking to label an enzyme.

The basis of the assay is that PerCp is in an activated form and I need only add the target protein and the conjugation will take place through free lysine residues. However, I don't see that...

28 January 2013 4,998 0 View