Dear All
a colleague claims that 1% SDS in lysis buffer is sufficient to inhibit proteases and there is no need to use protease inhibitors. is this correct?
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#2
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Posted 06 July 2009 - 11:28 AM
yobou, on Jul 6 2009, 07:56 PM, said:
Dear All
a colleague claims that 1% SDS in lysis buffer is sufficient to inhibit proteases and there is no need to use protease inhibitors. is this correct?
a colleague claims that 1% SDS in lysis buffer is sufficient to inhibit proteases and there is no need to use protease inhibitors. is this correct?
At least Proteinase K is not inhibited...
One must presume that long and short arguments contribute to the same end. - Epicurus
...except casandra's that belong to the funniest, most interesting and imaginative (or over-imaginative?) ones, I suppose.
#3
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Hmmm, I think it's working
Posted 06 July 2009 - 04:25 PM
It is false - SDS is a detergent, it primarily dissolves lipids rather than proteins.
Edited by bob1, 06 July 2009 - 04:26 PM.
#5
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Posted 07 July 2009 - 07:28 AM
HomeBrew, on Jul 7 2009, 12:23 PM, said:
Proteinase K's activity is actually enhanced by SDS: "Activity can be stimulated by addition of denaturing agents (SDS and urea)" (see here, under "Activators").
I mean proteases in human cancer cells (serine, cysteine and aaspartate , ...etc proteaes) . I guess proteinaseK is not among them.
#6
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Posted 07 July 2009 - 02:48 PM
Proteinase K is a serine protease from a eukaryotic organism...
Regardless, in answer to your question -- I don't think 1% SDS is effective enough to rely on it alone for protease inhibition.
Regardless, in answer to your question -- I don't think 1% SDS is effective enough to rely on it alone for protease inhibition.
#8
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Posted 08 July 2009 - 12:05 PM
GeorgeWolff, on Jul 8 2009, 06:36 PM, said:
Ask your colleague for the data.
actually before I open this topic for discussion I did so, and he showed me a blot for a protein that is clealry upregulated by certain drug treatment as a positive control. That is why I was asking whether this method is valid or not.
Edited by yobou, 08 July 2009 - 12:06 PM.
#9
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Posted 17 July 2009 - 04:22 AM
is there any reason you wish to avoid habitual protease inhibitors?
