where [ATP], [ADP], and [AMP] are the respective intracellu-
lar/tissue concentrations inμmol.4 Notes
- This is advised to remove microparticles that could block the
column and lead to increase in system pressure. - The hydrolysis of MgATP can take place very rapidly. EDTA is
therefore added to the extraction buffer for chelating magne-
sium in order to avoid ATP degradation by ATPases. - This is advised to remove microbubbles that could eventually
interfere with the UV spectrophotometer. - Pay attention to the direction of the column (seethe arrow on
the side of the column indicating the flow direction). - It is important to double-check that the volume of pyrophos-
phate buffer available is enough for the whole run.
Acknowledgments
Guigas’ group is supported by funding from the European Federa-
tion for the Study of Diabetes (EFSD/Lilly Research Grant Fel-
lowship), the Socie ́te ́Francophone du Diabe`te (SFD), and the
Dutch Organization for Scientific Research (ZonMW TOP Grant
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236 Noemı ́Garcı ́a-Tardo ́n and Bruno Guigas