Nitrite/Nitrate Determination Assay Kit

Details for Product No. ABIN3172710, Supplier: Log in to see
Detection Range
1-100 μM
Minimum Detection Limit
1 μM
Application
Biochemical Assay (BCA)
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Purpose The kit is designed for nitrite/nitrate determination in biological samples.
Sample Type Plasma
Detection Method Colorimetric
Specificity Specific for nitrite/nitrate determination in plasma samples.
Characteristics The overproduction of nitric oxide may lead to oxidative and nitrosative stress. It has been demonstrated that they enhance the development of a variety of diseases, as well as the ageing process1. Regarding nitrosative stress, high levels of iNOS have be
Components Plate and the reagents neccesary to perform the assay.
Material not included Pipettes, reaction tubes, plate reader.
Comment

For nitrite only: 50μL, For Nitrite/nitrate: 100μL

Assay Time 2 h
Plate Uncoated
Reagent Preparation

Reagent Preparation: 1. To prepare Reagent A solution: Add exactly 1000 μL of ultrapure water to Reagent A and mix thoroughly. This reagent must be freshly prepared. 2. To prepare Reagent C solution: Add exactly 1000 μL of ultrapure water to Reagent C and mix thoroughly. This reagent must be freshly prepared.

Sample Preparation

It is recommended to assay the samples in duplicate. Plasma samples may be deproteinized before performing the assay.

Assay Procedure

Standard Preparation: Prepare the calibrate in 1 mL tubes following the Table 1 (see kit booklet). Use ultrapure water as diluent. Performing the assay: The following procedure is for the determination of nitrite + nitrate: 1. Add 50 μL of the sample or standard in each well (96-well plate). 2.Add 10 μL of Reagent A and 20 μL of Reagent B. Incubate for 60 minutes. 3.Add 10 μL of Reagent C and 10 μL of Reagent D. Incubate for 20 minutes. 4.Add 50 μL of Reagent E in each well. Incubate for 10 minutes protected from light. 5.Add 50 μL of Reagent F in each well. Incubate for 10 minutes protected from light. 6.Read the absorbance at 540 nm within 30 minutes. In order to measure only the nitrite in the sample (not both nitrite and nitrate, "total nitrite"), add ultrapure water instead of Reagents A, B, C and D and continue the assay by adding Reagents E and F as shown in the procedure.

Calculation of Results

1.Calculate the average absorbance of each sample and control. 2.Determine the concentration in the sample by comparison to the Nitrite Standard reference curve (Figure 4 see Images). Nitrite (μM) = (?A540nm- intercept)/ slope

Restrictions For Research Use only
Preservative Sodium azide
Precaution of Use This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Storage -20 °C
Supplier Images
Biochemical Assay (BCA) image for Nitrite/Nitrate Determination Assay Kit (ABIN3172710) Typical standard curve