10981249001
Roche
Nitrate Reductase
from Aspergillus species
Synonym(s):
NAD(P)H:nitrate oxidoreductase
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form
lyophilized
specific activity
~0.4 units/mg protein (At 25 °C with nitrate as the substrate.)
packaging
pkg of 20 U
manufacturer/tradename
Roche
shipped in
wet ice
storage temp.
−20°C
General description
Nitrate reductase contains molybdenum as a cofactor and belongs to the family of dimethyl sulfoxide reductase. It is localized in periplasm, cytoplasm, and cell membrane.[1]
Application
- Nitrate Reductase is used for nitrate determination:Assay of nitrite and nitrate in culture media.
- Determination of NO3− in serum.
It was used to reduce nitrate to nitrite which will help in determination of nitric oxide metabolites levels in tissue using spectrophotometric methods.[2]
Nitrate reductase has been used to determine nitric oxide activity by measuring its stable metabolites, nitrate and nitrite upon reduction.[2]
Biochem/physiol Actions
Nitrate reductase is the first enzyme in nitrate assimilation pathway.[3] During reduction of nitrate to nitrite, it uses pyridinenucleotides, flavin, or benzyl viologen as electron donors.[4] It acts as central point for integration of metabolism by monitoring flux of reduced nitrogen in plants, algae and fungi.[5] In plants, nitrate reductase is regulated by a number of factors, such as nitrate, metabolites, dropdown in temperature, phytohormones, drought and light.[6]
Quality
Contaminants: <0.5% “NADPH oxidase”, <0.8% NAD(P)H-dependent ADH, <0.15% nitrite reductase
Preparation Note
Working concentration: Nitrate determination using Nitrate reductase:
The concentration of NADPH is 200 to 250 μmol/ml in the assay. This guarantees the function of the assay with regard to the stability of NADPH in the kit. This is the highest possible NADPH concentration for measuring an accurate E1.
Assay for the activity of Nitrate reductase:
Optimal NADPH concentration is 600 μmol/l and 5 μmol/l additional FAD is sufficient for the assay as the enzyme contains already FAD.
Storage conditions (working solution): A solution of 20 U Nitrate reductase in 2 ml double-dist. water is stable for one week when stored at 2 to 8 °C; for longer periods, freeze the solution in aliquots.
Nitrate Reductase is shipped at 15 to 25 °C.
Note: Avoid repeated thawing and freezing.
The concentration of NADPH is 200 to 250 μmol/ml in the assay. This guarantees the function of the assay with regard to the stability of NADPH in the kit. This is the highest possible NADPH concentration for measuring an accurate E1.
Assay for the activity of Nitrate reductase:
Optimal NADPH concentration is 600 μmol/l and 5 μmol/l additional FAD is sufficient for the assay as the enzyme contains already FAD.
Storage conditions (working solution): A solution of 20 U Nitrate reductase in 2 ml double-dist. water is stable for one week when stored at 2 to 8 °C; for longer periods, freeze the solution in aliquots.
Nitrate Reductase is shipped at 15 to 25 °C.
Note: Avoid repeated thawing and freezing.
Other Notes
For life science research only. Not for use in diagnostic procedures.
signalword
Danger
hcodes
Hazard Classifications
Resp. Sens. 1
Storage Class
11 - Combustible Solids
wgk_germany
WGK 2
flash_point_f
does not flash
flash_point_c
does not flash
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