Method	Sample Type	Sample Parameters	Information Content
Atomic Absorption Spectroscopy	Purified dioxygenase, purified reductase	Iron content 1–20 μM	Total iron content, correlated to protein concentration to estimate upper limit on complete active site content
UV-Visible Rieske Cluster Analysis	Purified dioxygenase	Protein concentration 3–10 μM	Qualitative Rieske cluster content reported as a ratio of Rieske peak absorbance intensity at 450 and 550 nm to protein absorbance at 280 nm
Steady State Activity	Purified dioxygenase, purified reductase	2 μM dioxygenase, 6 μM reductase	Activity following protein incubation at elevated temperatures relative to specific activity at temperature optimum
Lifetime	Purified dioxygenase, purified reductase	2 μM dioxygenase, 6 μM reductase	Total number of catalytic cycles each enzyme can undergo prior to loss of activity, typically irreversible; Here we outline real-time monitoring of NADH absorbance and discontinuous substrate quantification by HPLC.
DSC	Purified dioxygenase or reductase	20 μM protein	Heat capacity variation due to structural transition event as samples exposed to heat. The result of this method is a quantitative ΔH and mapping of the thermal dissociation pathway for the protein.