I have got 4-5 extra bands in SDS gel after running a my eluted protein which was purified through Ni-NTA column. This means impurities are present. My question is, can I again run my protein through Ni-NTA column to purify it perfectly? Will there be any problem regarding binding to the column when I will run it for second time?
Hi Subhecchha,
I suggest you try other purification methods to remove contamination bands. Ni-NTA resin may bind certain proteins non-specifically, thus it is common to have many protein bands after affinity chromatography. You need to perform further operations such as gel filtration or ion exchange to purify the protein of interest.
Hope it helps!
Roger Davis
Roger,
Thank you so much. I am planning to go for a Size exclusion. :)
- Badges
- Science topic
- Similar topics
- Biological Science
- Kinesiology
- Running