Could you please address this problem?
I am treating lung cancer cell line (A549) with Tetrahydrobiopterin (BH4) in DMSO. There is a slight effect, but BH4 is very unstable and it oxidizes very quickly (to BH2). Do you have any suggestions how to make the treatment more efficient? (better anti- oxidative conditions maybe?)
Thanks!
Hi,
Did tou try to add an antioxydant to your treatment like ascorbic acid or polyphenols ?
Sincerely
Dear Bertrand,
Thank you for your suggestion!
No, we haven't tried with antioxidant. How would you suggest me to add ascorbic acid for example- directly to the media or into the BH4 stock? Do you know what is the recommended concentration?
Thanks
Hi,
Honestly, it's difficult to propose anything. I think you will have to try various combinations, to determine what is the best solution.
In my opinion, it's best to add the antioxydant to the BH4 solution. About the concentration, you will have to test what is the concentration that best protect your BH4 solution without harming its activity on cells.
I hope it helps
Sincerely
Hi there,
Use sepiapterin, which easily enters the cell. BH4 has a cycle in the membrane and just a percentage will be uptake.
Also, any pH higher than 4 will facilitate the transformation of 98% oh BH4 into BH2. So prepare the stock under acidic conditions plus DTE 6.5mM
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