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Activity of Acetyl-CoA Carboxylase or
Malonyl-CoA Decarboxylase in Tissues
   
• For acetyl-CoA carboxylase, the tissue extract is incubated with unlabeled acetyl-CoA + NaH13CO3 to form [13C]malonyl-CoA which, after hydrolysis, is assayed as its TMS derivative.

[U-13C3]Malonyl-CoA is used as an internal standard. 

• For malonyl-CoA decarboxylase, we developed two different assays. The first assay involves incubation with [2-14C]malonyl-CoA, to form [2-14C]acetyl-CoA which is reacted with carnitine to form [2-14C]acetyl-carnitine the radioactivity of which is counted after isolation.

• The second assay involves incubation with [U-13C3]malonyl-CoA to form [1,2-13C2]acetyl-CoA.  The latter is reacted with thiophenol, and acetylthiophenol is assayed by GC-MS, [2H3, 1-13C]acetyl-CoA is used as internal standard.