Air during sample loading
when I was applying a cell lysate to a FPLC column, air bubbles were accidently introduced from the bottom. Is there any possibility that the amino acids got modified (e.g. oxidation of cysteins (although 3 mM BME was present in the solution))? The proteins in the lysate were denatured as it contained 6 M guanidine hydrochloride. So I am only concerned about chemical modifications.
Re: Air during sample loading
I don't think you have to worry about amino acids getting modified (unless your wanting too look at reduced/oxidised condition of your protein e.g. DTT for reduced conditions). This has happened to me also, the air might affect binding of your column if it gets stuck.
Last edited by Kneeanderthal; 10-22-2012 at 07:19 PM.
|air , loading , sample|
|Thread||Thread Starter||Forum||Replies||Last Post|
|SDS-PAGE Protocol||moleculardude||Protein Science||14||02-12-2010 10:27 AM|
|No Bands on Western Blot||clokke76||Western Blot Forum||2||08-27-2007 04:26 PM|
|SDS precipitates when I add my sample to PAGE sample email@example.com||Protocols and Methods Forum||6||06-18-2006 11:25 PM|
|Sample Preparation Workshop - Ottawa - November 16, 1st Announcement.||Victor Benham||Forum Chemie||0||11-05-2004 12:56 AM|
|Damped Harmonic Oscillation (new thread)||Anselm Proschniewski||Forum Physik||19||12-19-2003 10:29 AM|