We want to measure protein synthesis and degradation in live cultured tissue or cell samples and also zebrafish larvae (mg quantities) using tritiated water (as one can with D2O in MS). I have done this with S35-met in retic lysates in the 1980s, but I am sure things have moved on in terms of LSCs, scintillants, solvation etc. Can anyone recommend a simple protocol, or where to start looking?
Hello Simon,
sorry to see that your very interesting technical question has not yet received any expert answers. As a synthetic inorganic chemist I would not call myself a proven expert in this field. Although we have some experience with radioactive compounds (especially uranium and thorium) we never worked with tritium-labeled compounds. However, I just came across a potentially useful article in which a protocol for quantifying tritiated amino acids in proteins is described:
A sensitive method for measuring protein turnover based on the measurement of 2-3H-labelled amino acids in protein
This paper is freely available as public full text (see attached pdf file).
Good luck with your work and best wishes!
- Badges
- Science topic
- Similar topics
- Phytochemistry
- Extracts