Cell-based antioxidant protection assay
AssigneeNatural Immune Systems, Inc.
InventorGitte S. Jensen
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Abstract
Methods are provided herein for determining antioxidant activity of a test sample in intact cells. The method includes determining the antioxidant capacity of a test sample in intact red blood cells, wherein the test sample is added to intact red blood cells and oxidative damage is measured by alteration of fluorescence intensity of an oxidation-sensitive fluorescent indicator dye.
Claims20
A method for determining antioxidant activity of a sample in red blood cells comprising a fluorescent indicator dye which exhibits oxidation-sensitive changes in fluorescence intensity, the method comprising:
- adding a sample to intact red blood cells under conditions which permit the sample to cross the cell membrane of the intact red blood cells;
- washing the red blood cells to remove sample which has not crossed the cell membrane of the red blood cells;
- lysing the washed red blood cells;
- inducing oxidative stress in the lysed red blood cells;
- and
- detecting fluorescence intensity in the lysed red blood cells,
- wherein a decrease in fluorescence intensity in the lysed red blood cells in the presence of the sample as compared to a control indicates the antioxidant activity of the sample.
A method for determining antioxidant activity of a sample in red blood cells comprising a fluorescent indicator dye which exhibits oxidation-sensitive changes in fluorescence intensity, the method comprising:
- adding a sample to intact red blood cells under conditions which permit the sample to cross the cell membrane of the intact red blood cells;
- washing the red blood cells to remove sample which has not crossed the cell membrane of the red blood cells;
- lysing the washed red blood cells;
- and
- detecting fluorescence intensity in the lysed red blood cells,
- wherein a decrease in fluorescence intensity in the lysed red blood cells in the presence of the sample as compared to a control indicates the antioxidant activity of the sample.
Dependencies are inferred from the claim text.