In order to keep my Tris 20 mM GSH pH8 elution buffer active for as long as possible I would like to know if I could prepare a large quantity and freeze it? (how much time can I freeze/thaw it?) knowing that the half-life of reduced glutathione is short in aqueous solution and that preparing small volumes of buffer each time is not ideal… (weight a small amount of GSH)
Thanks !
Hi Dimitri
Check this PDF. I worked by purifying a GST enzyme from F. hepatica and I had prepare small aliquotes of elution buffer and stored at -20 degree.
Freezing will help against any microbial contamination, but does not necessarily help to keep reducing agents in their reduced state. You have to limit the exposure to air. Oxygen can diffuse faster in ice than water. You need to remove the oxygen in the headspace of your container; if your buffer is OK with it, bubble some N2 through it for a while.
I also found the following link:
https://www.researchgate.net/post/How_can_I_prevent_auto-oxidation_of_glutathione_in_my_sample
Dear Dimitry
i think that the simplest way is prepare a large volume of tris solution at ph8 with out dtt that can be stored at room temperature for long time and small aliquotes of concentrated gsh (eg 1M) that you can readly de-frozr and add to the tris buffer just before purification.
this is a standard Approach that yoy can use also for DTT for example.
ciao
Manuele
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