Hi All:
One of my crystallisation condition contains HEPES-Sodium. During the process of optimisation, I am trying to make HEPES-Sodium buffer using HEPES and NaOH. It will be really helpful if somebody can comment on it. I am also aware of the fact that there are several vendors from which we can buy. However, it is going to be my last option.
Thank You in advance.
Regards
Prasun
Hi Prasun, for example for a 1 M HEPES-NaOH prepare a 1 M HEPES solution (23.83 g / 100 ml) and adjust the pH with a 10 M NaOH solution to the pH that you need. Furthermore, I usually filter my buffer solutions (0.2 micrometer syringe filter) if I store them long. Also, I know the purity of HEPES can make a difference in crystallization so it may be worthwile to try HEPES from different vendors. Good luck, Tjaard
Hey Prasun,
How is it going there in Bristol?
Its fairly straightforward to prepare HEPES.NaOH. Dissolve the required amount of HEPES into whatever volume you want to prepare, the pH would be below 7 as HEPES itself is a sulfonic acid based small molecule. Adjust the pH by using a concentrated solution of NaOH or even you can use NaOH beads in case you are preparing larger volumes. Thats it! I use HEPES.KOH for many of my SEC and assays etc ans that's how I prepare it.
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