Hi All,
Has anyone got experience or know of a way to separate soluble protein from coacervates that are formed by the charge balanced between the protein and RNA?
This needs to be a way that will allow me to complete CD on the soluble phase and the coacervates.
I have attempted normal centrifugation but with no success. Do we think that ultracentrifugation would be too disruptive?
Many thanks,
My suggestion is to use gel filtration chromatography (by gravity) using a high-porosity resin such as Bio-gel A0.5m or A5m, depending on the size of the coacervates. Being much larger than the original protein, they will elute first.
Adam B Shapiro Thanks for the suggestion! I hadn't considered doing this as I haven't got much experience in this. I will do some research and see what I find.
Thank you very much!
Protein purification and separation is a series of processes intended to isolate one or a few proteins from a complex mixture. Separation of one protein from all others is typically the most laborious aspect of protein purification. Separation steps usually exploit differences in protein size, physico-chemical properties, binding affinity and biological activity. It is difficult to answer shortly. For soluble protein you may first follow salting out method followed by centrifugation and dialysis. but it depend on your purpose. For basic information you may follow link: http://www.aquaculture.ugent.be/Education/coursematerial/online%20courses/ATA/analysis/prot_sep.htm
and article for appropriate methods.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Nucleic Acids
- RNA