Product Name:
Nitric Oxide Assay Kit
Description:
The Nitric Oxide Assay Kit determines nitric oxide composition through measurement of nitrate (NO3) and nitrite (NO2) levels in urine, saliva, plasma, serum, and other biological fluids.
Application:
Functional Assay
Advantages:
Assay Kit containing the folllowing: Assay Buffer, Enzyme Cofactor, Enhancer, Nitrate Reductase, Nitrate Standard, Nitrite Standard, DAN Probe, Sodium Hydroxide, Microtiter plate, Plate Cover
Detection Principle:
Two microplates are necessary to perform this assay. One plate is used to measure the amount of nitrite in each sample. For the second plate, all of the nitrate in each sample is converted to nitrite using nitrate reductase enzyme and the “total” amount of nitrite (pre-existing nitrite plus nitrate that was converted) is measured. The nitrite measurements from plate 1 are subtracted from the total nitrite measurements in plate 2 in order to calculate nitrate concentrations. This kit is specifically designed for researchers concerned with the relative levels of nitrate and nitrite in each sample.
Format:
Colorimetric stain (azo dye) for clear 96-well plates
Intended Use:
Assay Kit for activity of Nitric Oxide in the research laboratory
InterAssay Precision:
Dependent upon several factors (generally, intra- and inter-assay CV ≤ 5%)
Introduction:
Nitric oxide (NO) plays an important role in neurotransmission, vascular regulation, immune response and apoptosis. Since NO is rapidly converted to nitrite and nitrate, the total concentration of nitrite and nitrate is used as a quantitative measure of NO production. The Nitric Oxide Fluorometric Assay Kit provides an accurate and convenient measurement of total nitrate/nitrite concentration in a simple two-step process. In the first step nitrate is converted to nitrite by nitrate reductase. In the second step, nitrite reacts with the fluorescent probe DAN (2, 3 diaminonaphthalene). NaOH enhances the fluorescent yield. The fluorescent intensity is proportional to the total nitric oxide production. The kit has been tested with culture media, plasma, and tissue homogenates.
Kit Contents:
Griess reagents, nitrite and nitrate standards, nitrate reductase, NADH, and diluents.
Measurement:
Absorbance at 540 nm (±20 nm)
Sample:
Urine, saliva, plasma, serum, and other biological fluids
Sensitivity:
0.222 μM nitrite; 0.625 μM nitrate
Storage Conditions:
Store at -20°C.
Testing Time:
15 minutes (nitrite assay); 45 minutes (nitrate assay)
