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Protein degradation


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#1 MariaG

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Posted 18 March 2009 - 06:19 AM

Dear all,
I have yet another problem with the protein I am purifying, I have tested the quality and amount of purification on SDS-PAGE followed by coommassie staining, there was a very nice and clear band, it was a month ago. Today I have done SDS-PAGE and coommassie again and the same band of the same protein and the same amount of the protein is barely seen! Therefore I have concluded that the protein was partially degraded. I keep it at 4 C, since it looses activity if frozen, I sterilize it with 0.2 uM filter.
Is there something I can do in order to prevent protein degradation? Add glycerol? Increase salt concentration?
Thank you very much in advance.

#2 madrius1

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Posted 18 March 2009 - 09:07 AM

Dear all,
I have yet another problem with the protein I am purifying, I have tested the quality and amount of purification on SDS-PAGE followed by coommassie staining, there was a very nice and clear band, it was a month ago. Today I have done SDS-PAGE and coommassie again and the same band of the same protein and the same amount of the protein is barely seen! Therefore I have concluded that the protein was partially degraded. I keep it at 4 C, since it looses activity if frozen, I sterilize it with 0.2 uM filter.
Is there something I can do in order to prevent protein degradation? Add glycerol? Increase salt concentration?
Thank you very much in advance.


What about proteases inhitibors?

#3 mdfenko

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Posted 18 March 2009 - 11:32 AM

if you can't freeze it then you can try adding glycerol to about 50% and then store at -20C. it won't freeze but should survive longer.
talent does what it can
genius does what it must
i do what i get paid to do




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