Yellow Laemmli Sample Buffer - (Apr/02/2008 )
ahh, TCA precipitation, that makes sense. So the pH probably isn't too bad for the protein, just as long as I don't see precipitation, right?
-MKR-
QUOTE (MKR @ Apr 6 2008, 01:16 PM)
ahh, TCA precipitation, that makes sense. So the pH probably isn't too bad for the protein, just as long as I don't see precipitation, right?
in principle yes, if you use organic acids; anorganic acids should be avoided to precipitate proteins for analysis
-The Bearer-
QUOTE (MKR @ Apr 5 2008, 03:37 AM)
I want to know why you would throw it out. What is bad about such a buffer?
So, to clarify, the question is - what would the low pH do to the protein samples?
i would throw out the buffer because usually it is only a small amount and would otherwise have to calculate amounts of additional components to add for the expected increase in volume.
if the pH is low enough then you may neutralize the negative charge of the sds and alter the migration of the sample.
-mdfenko-
>>neutralize the negative charge of the sds and alter the migration
I had not thoght about this. Thanks.
-MKR-
As said The Bearer and mdfenko, you should throw away your sample buffer (I didn't know if you had already your samples in the sample buffer).
there are several known reasons for your experiment not to work, only because the pH is wrong. It would be better to prepare new sample buffer than to get a chance that your experiment doesn't work.
-Missele-
QUOTE (Missele @ Apr 9 2008, 05:21 AM)
As said The Bearer and mdfenko, you should throw away your sample buffer (I didn't know if you had already your samples in the sample buffer).
there are several known reasons for your experiment not to work, only because the pH is wrong. It would be better to prepare new sample buffer than to get a chance that your experiment doesn't work.
there are several known reasons for your experiment not to work, only because the pH is wrong. It would be better to prepare new sample buffer than to get a chance that your experiment doesn't work.
What are those reasons?
-MKR-