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Inclusion bodies in e.coli


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#1 bhavana

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Posted 11 September 2010 - 08:57 AM

how to process further with inclusion bodies for loading in SDS-PAGE to see presence of protein, what is the lysis buffer i should use, when i used lammeli buffer and heat the inclusion bodies i observed formation of very tough pellet which i can not able to load it in the SDS gel, cn anybody plzzzz help me out........

#2 mdfenko

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Posted 17 September 2010 - 10:01 AM

instead of boiling you can heat at 60-70C for 10-20 minutes. boiling can cause aggregation of the protein despite the sds and 2-me. but be careful that you don't use too much protein, you won't get it all into solution.
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