The OxiSelect Ferric Reducing Antioxidant Power (FRAP) Assay Kit is a quantitative assay for measuring the antioxidant potential within a sample. Ferric iron (Fe3+ ) is initially reduced by electron-donating antioxidants present within the sample to its ferrous form (Fe2+). The iron colori-metric probe complex develops a dark blue color product upon reduction which can be measured at 540-600 nm (see Figure 1). Samples can be compared to the iron standard for determining antioxidant potential. This assay is analytically sensitive to approximately 4 µM of Fe2+ iron equivalents, or a FRAP value of 2.
Storage:
Upon receipt, store the kit components at 4ºC.
1X Assay Buffer:
Prepare 1X Assay Buffer by adding deionized water to the 5X Assay Buffer (e.g. add 3 mL of 5X Assay Buffer to 12 mL of deionized water). Mix thoroughly until homogeneous. Use this buffer for preparing kit reagents and within the assay. Store at 4ºC when not in use.
Reaction Reagent:
Prepare the Reaction Reagent just before use and prepare only enough for immediate applications. Dilute the Colorimetric Probe 1:10 and the Iron Chloride Solution 1:10 in 1X Assay Buffer (e.g. For 50 assays, add 500 µL Colorimetric Probe and 500 µL Iron Chloride Solution to 4 mL 1X Assay Buffer for 5 mL total). Vortex thoroughly.
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