Could denatured proteins be stored at -20°C (or -80°C) until use (for SDS-PAGE)?
Posted 23 April 2009 - 08:11 AM
Mix the sample with equal volume of 2X loading/sample buffer; boil the mixture (denaturation) at 95 °C for 3 min; cool down the samples on ice for 5 min; store the samples at -20°C (or -80°C) until use.
I actually followed this protocol and had no problem to detect protein expression with Western blot. However, I am not sure if this is a good idea. Is there any literature support?
Posted 23 April 2009 - 11:47 AM
I used protein samples for western blot which were stored that way for more than one year (-20°C), and everything was fine!
Posted 23 April 2009 - 12:13 PM
It's generally accepted that proteins under very cold conditions are more or less in stasis (most things are). If possible, I'd also store them in a medium which will freeze at -80*C, such that the protein molecules will be sterically hindered by the frozen solvent. Keep them as cold as possible just to be safe. ... That would be my advice.
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