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SAM and SAH quantification

CG Carolina Magdalen Greco

Last updated date: Mar 1, 2021 Views: 818 Forks: 0

original An abbreviated version of this protocol was published in Science Advances in Dec, 2020
S-adenosyl-l-homocysteine hydrolase links methionine metabolism to the circadian clock and chromatin remodeling
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  1. Plate MEF cells in 10 cm petri dishes
  2. Once confluent, treat MEF cells with vehicle (DMSO) or deazaneplanocin A (DZnep) for 24 hours
  3. Scrape cells in cold PBS and collect by centrifuging 1,200 rpm for 10 minutes at 4 °C
  4. Resuspend cells in 200 ml cold PBS
  5. Sonicate on ice
  6. Centrifuge at 14,000 rpm for 15 minutes at 4 °C
  7. Remove the supernatant and store on ice
  8. Determine SAM and SAH levels using 50 ml of supernatant according to the manufacturer’s instructions (STA-671-C, Cell Biolabs).

 

Solutions:

 

Phosphate-buffered saline (PBS)

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How to cite：
Readers should cite both the Bio-protocol preprint and the original research article where this protocol was used:
  1. Greco, C(2021). SAM and SAH quantification. Bio-protocol Preprint. bio-protocol.org/prep890.
  2. Greco, C. M., Cervantes, M., Fustin, J., Ito, K., Ceglia, N., Samad, M., Shi, J., Koronowski, K. B., Forne, I., Ranjit, S., Gaucher, J., Kinouchi, K., Kojima, R., Gratton, E., Li, W., Baldi, P., Imhof, A., Okamura, H. and Sassone-Corsi, P.(2020). S-adenosyl-l-homocysteine hydrolase links methionine metabolism to the circadian clock and chromatin remodeling . Science Advances 6(51). DOI: 10.1126/sciadv.abc5629

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