A radiochemical assay for heme synthase (ferrochelatase) activity is described in
this report. The principle of the assay is to measure the incorporation of 59Fe, added to the
reaction flask with a known amount of iron, into heme. Iron is maintained in the ferrous state,
and oxygen is excluded from the flask during the incubation of the substrates with enzyme.
Labeled heme product is extracted by solvent partitioning and counted in a gamma scintillation
spectrometer. The assay can detect picomole amounts of product and is most useful when
low levels of heme synthase activity are present.
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