Hi all,
Is there a way to extract RNA and protein from the same cell culture sample? Because we need to use the protein for gelatin zymography at later time, we are looking for a method that can obtain high quality RNA, including miRNA, and un-denatured protein, from the same sample. The Trizol method, which denatured the protein according to manual, seems not appropriate. Please let me know if that's not the case.
Thank you all.
I also would like to know if such method exists.
The main issue here is that you need to extract proteins/enzymes and keep them in their native active form to perform the zymograms.
Could you split the cells into 2 parts, one for DNA RNA etc. the other part for protein.
I've read about such a method, although I've never performed this procedure myself.
There's a kit called PARIS from ThermoFisher; link below. Could it be something you're looking for?
https://www.thermofisher.com/pl/en/home/references/ambion-tech-support/rna-isolation/tech-notes/a-better-way-to-isolate-rna--and-protein-from-the-same-sample.html
Generally, native proteins and RNA are only in the cell. Most proteins and RNA in the cell are in the form of protein-protein and RNA-RNA or RNA-protein complexes. It is necessary to destroy these complexes to isolate the target RNA and target protein. It disrupts their native structure. However, the structure of proteins (not high-molecular and not subunit) can be restored to some degree under conditions close to cellular ones. The easiest way to look at the renaturation of enzyme proteins is to restore their activity. If any other criterion of the native protein is known for the target protein, one can restore its nativeness.
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