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Air during sample loading

Air during sample loading - Protein Forum

Air during sample loading - Protein Forum


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Old 09-01-2012, 03:34 PM
Pipette Filler
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Default Air during sample loading



Hi,

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.

Thanks.
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Old 10-22-2012, 07:18 PM
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Default 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.
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