One of the methodologies I would like to use state that they adjusted the pH of their protein solution to 9. However, they did not state how they did this. I am wondering if I should go ahead and use 0.1M NaOH to achieve this or should I use a buffer? My concern about using a buffer is I might end up diluting the initial solution too much already. And my concern with using the base is that this might not be acceptable (too basic). Any recommendations regarding this would be really helpful. Thank you.
With the NaOH you risk you will precipitate your protein (local alkaline pH). If you'd use e.g. 1M Tris (either without adjusted pH or e.g. pH 10), you should not dilute your sample too much. What buffer do you use (the one you want to change pH of)?
You can always try your buffer without the proteins to see how much of the solution you'd need.
Thank you Thomas for the point about local pH increase and precipitation. the protein is not in any water, it is just diluted in water right now. Thank you for the tip, I'll do that checking first
It is in some buffer, isn't it? What the buffer is and what is the concentration?
sorry, I meant the protein is not in any buffer*. It is merely dissolved in water.
I see. Is that wise? Wouldn't it be good to use at least 5-20 mM buffer?
Anyway, if you have it only in water, the Tris pH 9.0 will work perfectly (basically just dilution of stock buffer, except instead of water you'll use your protein solution).
use a desalting column and exchange the buffer. that would be the fastest way to do it. dialysis is always risky.
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