I have used STAT60 to isolate RNA successfully. I am wanting to collect RNA and protein from tissues, and according to the manufacturer's information I could allegedly use STAT60 to collect both types of molecules. What I would ideally like to do is precipitate both RNA and protein from the same tissues and submit the protein for mass spectrometry. Has anyone used STAT60-isolated proteins successfully for mass spectrometry?
Hi Katherine,
the question is what you are looking at. Are you going to perform MS for global proteomics analysis via digestion (InGel or InSolution)and LC-MS of the peptides or pergorm MALDI-MS to identify single proteins after InGel digestion?
However be aware that in the STAT60 procedure precipitation of proteins is included which can result in different recovery of proteins between samples to compare and also can lead to a bias towards proteins which are precipitating easier with the used technique and therefore possibly lead to a loss of proteins which are not precipitating iso well.
You also need to consider that if you want to perform an InSolution digestion of the precipitated proteins for global LC-MS based proteomics you need to resuspend the protein pellet in an appropriate buffer or if you want to use the solubilisation buffer of 1% SDS, 10M Urea which is proposed in the STAT60 protocol (At this point I am wondering how they are able to resolve 10M Urea without heating?), You can use the FASP (Filter Aided Sample preparation method for the digestion.of your proteins.
Good Luck
Murat
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