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Protein Purification-catastrophe while purifying?


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2 replies to this topic

#1 joeysmi

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Posted 12 September 2013 - 10:24 AM

Hey forum

 

I just added fresh bacterial culture to a previously pellet that was stored overnight at -20C and spun down the bottle to get a larger pellet. Question is whether the bacteria at -20C was already lysed and I lost my protein from the first pellet when I threw out the second supernatant? thanks for your advice.



#2 tom43

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Posted 12 September 2013 - 11:23 AM

Depends on cells and protein, but: Freezing is an effective lysis method. If the frozen pellet was thawed by the second suspension you probably washed out the protein.



#3 labtastic

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Posted 25 September 2013 - 05:15 AM

You probably lost some protein having lysed the cells in your first pellet by freezing at -20C, but the loss shouldn't be too dramatic. I would still go ahead with the purification.






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