Again, this is regarding to my cloning work. I clone my protein in pQE-30 vector and tranfect into E.Coli (Qiagen product). After induced by IPTG, the bacteria was harvested and protein of interest was purified using Ni-NTA method acroding to manufacturer instruction.
However when I run SDS-PAGE, I still can see a lot of my protein in the cell lysate. My protein is a soluble protein, why it can be mixed with cell lysate??
From MERCK website I got this information that: "soluble protein has to be span at 10,000g at least 30 min before it can be seperated from insoluble protein/homogenized in supernatant."
Is that true?
I will be glad if someone can comman on this
Thank everybody : )
Submit your paper to J Biol Methods today!
1 reply to this topic