I am trying to make NADPH solution but because of its hygroscopic nature, it has absorbed water and it is hard to mass it out. My current plan is to just dissolve it in 10 mM NaOH and quantify it at A340 then store it at -80. It seems the compound is stable at these conditions for longer periods. Thanks!
Dear Erome,
maybe you could try to dissolve it in TRIS-HCl buffer (pH 8),even DMSO, or methanol.. Think that it could be stored for a few weeks up to 6 months at -80 degrees C.
Good luck!
If the solid has turned yellow(ish), I would fear some oxidation to NADP+ has occurred as well, which may obscure your enzyme studies. Regardless of your storage method, I would double check NADPH concentrations through a known enzyme assay where the michaelis-menten parameters are well-established in that system to ensure co-factor integrity.
Thank you both for your response. David, thanks for bringing that up. I am looking into some assays that I can do to establish the exact NADPH concentration by taking the oxidation into account.
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