Nitric Oxide (NO) Colorimetric Assay Kit

    • Biochemical Assay Kit-Elabscience
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    >
    • Biochemical Assay Kit-Elabscience
    • Biochemical Assay Kit-Elabscience

      Catalog number:E-BC-K035-M

      Size:
      • 48T
      • 96T
      Qty:
      - +
      Price: $120

      Detection method: Colorimetric method

      Detection instrument: Microplate reader

      Valid period: 6 months

      Lead Time: 7~10 daysWelcome to order from local distributors.

      Add to cart Compare Bulk request Manual

      General information

      Detection significance

      The half-life of NO is extremely short, it exists in form of nitrate or nitrite produced by vascular endothelial cell, vascular smooth muscle cell, platelet, and macrophage and so on. The concentration of NO can be indirectly measured by detecting that of nitrate or nitrite.

      NO react with oxygen and water to generate nitrate or nitrite which can form a kind of pale red azo compound when meet with nitrate chromogenic reagent, the absorbance of the compound can be measured to calculate the concentration of NO indirectly.

      Detection principle

      NO is easily oxidized to form NO2- in vivo or in aqueous solution, and a reddish azo compound is formed with the color developing agent, and the concentration of the azo compound is linearly related to the concentration of NO. The concentration of NO can be calculated indirectly by measuring the OD value at 550 nm.

      The key point

      1.     Use disposable EP tubes or clean glass tubes with stopper for centrifugation.

      2.     The supernatant for assay should not contain sediment, otherwise it will affect the results.

      3.     Samples can be stored at -20 for 1-2 months, the lower the temperature, the longer the storage time.

      4.     All reagents should be prepared the day before the assay, let it fully dissolved. Please add reagents to the bottom of well vertically and slowly, avoid to add on the wall of well and    generate bubble. 

      Operation procedures

      Plate set up

       

      1

      2

      3

      4

      5

      6

      7

      8

      9

      10

      11

      12

      A

      A

      A

      S1

      S9

      S17

      S25

      S33

      S41

      S49

      S57

      S65

      S73

      B

      B

      B

      S2

      S10

      S18

      S26

      S34

      S42

      S50

      S58

      S66

      S74

      C

      C

      C

      S3

      S11

      S19

      S27

      S35

      S43

      S51

      S59

      S67

      S75

      D

      D

      D

      S4

      S12

      S20

      S28

      S36

      S44

      S52

      S60

      S68

      S76

      E

      E

      E

      S5

      S13

      S21

      S29

      S37

      S45

      S53

      S61

      S69

      S77

      F

      F

      F

      S6

      S14

      S22

      S30

      S38

      S46

      S54

      S62

      S70

      S78

      G

      G

      G

      S7

      S15

      S23

      S31

      S39

      S47

      S55

      S63

      S71

      S79

      H

      H

      H

      S8

      S16

      S24

      S32

      S40

      S48

      S56

      S64

      S72

      S80

      Note: A: blank wells; B-H: standard wells; S1-S80: sample wells


      The dilution of standard curve

      Dilute 2 mmol/L sodium nitrite standard solution with deionized water to a serial concentration. The recommended dilution gradient is as follows: 300, 200, 150, 100, 40, 20, 10, 0 μmol/L

      Operation table

      1.       Pre-treatment

       

      Standard tube

      Sample tube

      Different concentrations of sodium nitrite (μL)

      A*

       

      Sample (μL)

       

      A*

      Reagent 1 (μL)

      200

      200

      Reagent 2 (μL)

      100

      100

      Mix fully and stand for 15 min at room temperature, centrifuge it at 3100 g for 10 min, take 160 μL of the supernatant for the following procedure.

      Note:  a: For serum or plasma samples, A* is 200-300 μL.

                b: For tissue or cell homogenates, A* is 100-300 μL.

      2.       Reaction

       

      Standard tube

      Sample tube

      Supernatant (μL)

      160

      160

      Chromogenic reagent (μL)

      80

      80

      Mix thoroughly for 2 min, stand for 15 min at room temperature, measure the OD of each well with micro-plate reader immediately at 550 nm wavelength.

       

      Performance characteristics

      Technical parameter

      Detection range 0.16-300 μmol/L Average inter-assay CV 3.7%
      Sensitivity 0.16 μmol/L Average intra-assay CV 2.4%
      Average recovery rate 102%

      Citations

      1. Publication: SONG Quan Quan, NIU Jing Ping, ZHANG Shu Yu, LIANG Ting Ting, ZHOU Ji, FENG Shan Shan. Effects of Simulated Heat Wave and Ozone on High Fat Diet ApoE Deficient Mice. Biomedical and Environmental Sciences, 2018, 31(10): 757-768.
        Species: Mouse
        Sample Type: Plasma and Heart homogenate
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