I am trying to generate nuclear extracts from tissue. Most of the buffers for this protocol contain HEPES, and I have seen a variety of pHs listed: 7.6, 7.9, and 8.0, with 7.9 being by far the most common. Does anyone know how sensitive this protocol is to pH and how much variability it will tolerate?
I ask because I just made HEPES, aiming for pH 7.9 and resulting in pH 7.95, and I'm wondering if that's ok. If not, what acid should I use to adjst the pH down? I used KOH to increase it.
Also, if anyone has insight into why pH 7.9 is the most desired, I would like to know.
nuclear extract -- HEPES pH?!
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