Description
• Direct, rapid quantitation of intracellular nitric oxide in cultured cells
• Detects nitric oxide as low as 3 nM
• Fluorometric probe is nitric oxide-specific and has excellent photostability and pH stability
• Suitable for flow cytometry, fluorescent microscopy, and fluorometric microplate detection
The OxiSelect Intracellular Nitric Oxide Assay Kit is a cell-based assay for rapid detection of intracellular NO or NOS activity in cultured cells. In brief, the cell-permeant NO probe passively diffuses into cells and is deacetylated by cellular esterases to a non-fluorescent intermediate. When intracellular nitric oxide encounters this intermediate, it rapidly oxidizes to a highly fluorescent, triazolo-fluorescein analog. The fluorescence intensity is proportional to the NO levels within the cell cytosol (detection limit of ~3 nM). The kit's provided Nitric Oxide Fluorometric Probe is NO specific, has excellent photostablity and pH stability, and is detected with standard fluorescein filters. The probe is suitable for flow cytometry, fluorescent microscopy, and fluorescent microplate detection. Each kit provides sufficient reagents to perform up to 96 assays, including blanks, standards, and unknown samples.
Introduction: Reactive nitrogen species (RNS) and reactive oxygen species (ROS) work together to damage cells and have been implicated in the pathogenesis of several disease states. RNS are a family of molecules derived from nitric oxide (NO) and superoxide anion (O2-), produced via nitric oxide synthase (NOS) and NADPH oxidase, respectively (Figure 1). Nitric oxide is an established mediator in vascular diseases, diabetes, renal ischemia, atherosclerosis, inflammatory diseases, and cancer. However, because of its extremely short half life, studies of NO and its physiological role have been challenging. The OxiSelect Intracellular Nitric Oxide Assay Kit is a cell-based assay for rapid detection of intracellular NO or NOS activity in cultured cells. In brief, the cell-permeant NO probe passively diffuses into cells and is deacetylated by cellular esterases to a non-fluorescent intermediate. When intracellular nitric oxide encounters this intermediate, it rapidly oxidizes to a highly fluorescent, triazolo-fluorescein analog. The fluorescence intensity is proportional to the NO levels within the cell cytosol (detection limit of ~3 nM). The kit's provided Nitric Oxide Fluorometric Probe is NO specific, has excellent photostablity and pH stability, and is detected with standard fluorescein filters. The probe is suitable for flow cytometry, fluorescent microscopy, and fluorescent microplate detection. Each kit provides sufficient reagents to perform up to 96 assays, including blanks, standards, and unknown samples