This may be a silly question, but is it the final pH of the...

Dec 14, 2012
protocol Protocol: Purification of His-ubiquitin Proteins from Mammalian Cells
This may be a silly question, but is it the final pH of the solution that varies between buffers, or only the pH of the TRIS used?

Thanks much!
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Vanesa Olivares-Illana Author Answered Jan 28, 2013

Instituto de Física, Universidad Autónoma de San Luis Potosí

Hello and sorry for the delay in the answer.
Im agree with Lin, is commonly accepted that the pH is the Tris pH. Just one point, in the case of wash buffer you have also the component of phosphate buffer around 8, but you slowly will go down the pH (because of the explanation of Lin) of each buffer to improve the purification.
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Lin Fang Answered Dec 22, 2012

Department of Pediatrics, School of Medicine, Stanford University, USA

I am not the author, but let me try to answer your question: I would think it is commonly accepted that the pH is the Tris pH. As a result, the buffer ends up in the same (or similar) pH as Tris. But it is a good lab practice to always check pH of the buffer especially when it is your first time to make such buffer. Another caution is that samples you will add in the buffer does not maximize its buffer capacity, which is also done through sensibly checking pH.
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