Glutaraldehyde works best at low pH
On Mon, Nov 7, 2011 at 8:40 AM, Ed Pozharski <[log in to unmask]> wrote:
> On Mon, 2011-11-07 at 05:19 +0000, Sam Arnosti wrote:
>> Hi everyone
>>
>> I have a protein that is extraordinarily stable at PH=3.0 or even 2.0.
>>
>> I want to crystallize it in the low PH and compare the differences between the crystals in regular PH and low PH.
>>
>> I was wondering how people set up the boxes in low PH, as usual buffers are mostly less acidic.
>>
>> Regards
>>
>> Sam
>
> Not clear if you already have crystals at "regular pH", but if you do,
> you may consider direct transfer to lower pH. Of course, crystals may
> dissolve, which you could possibly prevent by cross-linking with
> glutaraldehyde. Three caveats:
> a) If lattice is incompatible with lower pH, even with cross-linking the
> resolution may sink to essentially useless levels
> b) I have no idea if the cross-linking will not be disrupted at really
> low pH, perhaps someone else can comment on that
> c) the 3rd reviewer can always say that lattice forces could have
> prevented a conformational change. But same goes for direct
> crystallization at low pH (but caries less weight).
>
> --
> "I'd jump in myself, if I weren't so good at whistling."
> Julian, King of Lemurs
>
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