Cell-based assay for detecting activation of poly (ADP-ribose) polymerase
AssigneeInotek Pharmaceuticals Corporation
InventorsCsaba Szabo, Laszlo Virag
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Abstract
The present invention relates to methods of detecting poly(ADP-ribose) polymerase (“PARP”) activity in cells and tissue by contacting a cell with an effective amount of biotinylated NAD+ and detecting the presence of incorporated biotin within the cell. The invention also relates to the identification oxidatively stressed tissues and cells by detecting PARP activity. The invention also relates to identifying a patient suffering from a disorder that causes PARP overexpression.
Claims20
A method for identifying a poly(ADP-ribose) polymerase-expressing cell, said method comprising:
- (a) contacting a cell with an effective amount of biotinylated NAD+, and
- (b) detecting the presence of incorporated biotin within the cell.
A method for identifying a poly(ADP-ribose) polymerase-expressing tissue, said method comprising:
- (a) contacting a tissue with an effective amount of biotinylated NAD+ , and
- (b) detecting the presence of incorporated biotin within the tissue.
A method for determining the amount of poly(ADP-ribose) polymerase activity in a cell, said method comprising:
- (a) contacting a poly(ADP-ribose) polymerase-expressing cell with an effective amount of biotinylated NAD+, and
- (b) determining the amount of incorporated biotin within the cell.
A method for identifying inflamed tissue, said method comprising:
- (a) contacting the tissue with an effective amount of biotinylated NAD+ , and
- (b) detecting the presence of incorporated biotin within the tissue.
A method for identifying a patient having an oxidative-stress disorder, comprising:
- (a) contacting a cell or tissue of the patient with an effective amount of biotinylated NAD+, and
- (b) detecting the presence of incorporated biotin within the cell or tissue.
A method for identifying a patient having a disorder, wherein the disorder is selected from a group consisting of inflammation, shock, reperfusion injury, stroke, myocardial ischemia, diabetes, diabetic endothelial dysfunction, atherogenesis, and atherosclerosis comprising:
- (a) contacting a cell or tissue of the patient with an effective amount of biotinylated NAD+, and
- (b) detecting the presence of incorporated biotin within the cell or tissue.
Dependencies are inferred from the claim text.