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. 2009 Feb;21(2):479–493. doi: 10.1105/tpc.108.059550

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Copyright © 2009, American Society of Plant Biologists

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Figure 7. — Peroxidase Activity of Maize Annexin Preparations Measured Using Amplex Red.

(A) Time course. Increase in fluorescence (arbitrary units [AU]) supported by 25 μg/mL annexin-enriched preparation at pH 7.4 in the presence of 1 mM H₂O₂ (closed circles) was abolished by heat inactivation (open circles).

(B) Time course of HRP activity (0.095 μg/mL) using conditions described in (A). Heat-inactivated HRP (open circles).

(C) Time course of fluorescence increase supported by 25 μg/mL highly purified ANN33/35 preparation at pH 7.4 in the presence of 1 mM H₂O₂ (closed circles); heat inactivated ANN33/35 preparation (open circles).

(D) Heme detection. Absorption spectrum shows peroxidase-competent annexin-enriched preparation (5 μg) has no Soret peak (∼410 nm) indicative of a heme group, in contrast with either 2.2 μM (7.3 ng) or 22 μM (73 ng) cytochrome C oxidase (CytC).