What is the advantages and disadvantages of using either PBST or TBST in western blotting, or while working with proteins in general? Are there other buffers which are also used for western blotting, or washing steps while working with proteins?

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    $\begingroup$ In my experience it actually doesn't matter which of these buffers are used for washing. I think this is much more a flavor of the respective lab. $\endgroup$ – Chris Jul 10 '16 at 17:10

I used to work for a company well-known for its modification state-specific antibodies, including phospho-specific ones, and they actually performed extensive in-house testing of PBST vs. TBST in Western blotting. Part of the reason the company chose to recommend the use of Tris-buffered saline over phosphate-buffered saline based buffers was the clearly-demonstrated fact that some phospho-specific antibodies just didn't work as well in PBST as they did in TBST, for whatever reason. As I recall (it's been quite a while since the testing was performed), some of the antibodies tested gave weaker signals, with more background, and one in particular just didn't work at all in PBST - no signal whatsoever. For that reason, the company chose to standardize its recommended Western blotting protocol with TBST, and it's been that way ever since.

That being said, I'm sure that the majority of antibodies out there would work just fine in either buffer. However, to be on the safe side, if a company states one buffer or other in their protocol, it would be best to use the recommended one.


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