I intend to dilute my protein samples in PBS but am concerned since this is a different buffer system compared to the Tris buffers used in PAGE. I would like to ask if suspending my proteins in PBS, aliquoting it, and mixing with electrophoresis sample buffer is just fine.
There is no harm in diluting protein samples in PBS and subsequently use for electrophoresis. But, it is ideal to use 50 mM Tris-HCl buffer, pH 7.5 for extraction of proteins and subsequent electrophoresis. PBS is a good extraction as well as solubilising buffer than Tris-HCl, we very often use PBS for extraction of proteins in tissue samples and electrophoresis under Tris-buffer system. However, it involves excellent working knowledge on your target proteins, type of tissues etc. to decide on appropriate buffers for extraction and electrophoresis.
I agree with Prof. Janarthanan Sundaram. You can go with TBS as the running buffer is of similar composition and it will facilitate faster separation of protein in gel.
Yes, it's fine. I do that and no problem at all. I also agree with the suggestions of others.
Yes, I use PBS in my samples for SDS PAGE.
In addition, I used 50 mM Tris-HCl buffer, pH 7.5 and it works very well.
And, I also agree with all suggestions of others.
Good job.
Best wishes.
Should i use Tris-citrate pH8 for homogenizing of protien sample for sds page instead of PBS.?
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