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Different centrifugation duration, different protein??


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#1 eaton

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Posted 19 September 2010 - 07:21 AM

Hi! everybody:
I use tissue lysis buffer to do protein extraction. The last step of protein extraction is centrifugation in 14000prm for 5 mins. If I increase the duration from 5 mins to 10 mins, will I get a different results, such as different protein? Or just make my protein purer? Thank you!

#2 protolder

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Posted 20 September 2010 - 09:29 PM

Hola, I think that the results were similar. This speed is enought to precipitate the majority of particulate fraction, and the soluble proteins will continue in the soluble fraction whatever was the time, but the best check is recentrifuge 10 min the supernatant of 5 min.In the second centrifugation you could be a very little or none pellet. It would be different if the first centrifugation was to a minor speed but you will be fractionating particulate fraction. Membranes, nucleus, and other cellular bodies. Good luck




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