Glucose oxidase test kit 100 tubes/48 samples

Measurement steps:

1. Preheat the enzyme-linked immunosorbent assay (ELISA) reader for at least 30 minutes and adjust the wavelength to 450nm.

2. Dilution of Reagent 3: Dilute Reagent 3 50 times with distilled water and mix as much as needed. Dilute reagent three with distilled water at a ratio of 1:49.

3. Preparation of working fluid: Add 100 μ l of reagent 2 to reagent 1 and mix thoroughly. The prepared reagent can be stored for one week at 4 ℃ in the dark. If there are few samples to be tested at once, reagent one and reagent two can be mixed and prepared in a ratio of 20ml: 0.1ml according to the actual dosage

4. Dissolve one bottle of reagent 4 in 5ml of distilled water (use up within one week after dissolution).

5. Sample determination (add the following reagents in sequence to a 96 well plate)

Special reminder: This product is only for scientific research use and should not be used for direct clinical testing on the human body.

Product Introduction:

Points to note when choosing anticoagulation:

① The amount of anticoagulant added to each sample should be consistent, and the amount of whole blood taken should also be as consistent as possible;

② After collecting anticoagulated whole blood, it is necessary to gently invert it and fully anticoagulate it to prevent some blood from not coming into contact with anticoagulants and causing coagulation;

③ A relatively large amount of plasma is collected from anticoagulated whole blood (0.4-0.5ml of plasma can be separated from 1ml of anticoagulated whole blood);

④ After the plasma collected for anticoagulation is frozen and stored, there may be flocculent turbidity when thawed. If there is any turbidity, it needs to be removed by centrifugation for measurement.

operating steps:

Before the experiment begins, all reagents should be equilibrated to room temperature (reagents cannot be directly placed at room temperature)Dissolve at 37 ℃); When diluting reagents or samples, they should be mixed thoroughly, and foaming should be avoided as much as possible during mixing. Before the experiment, the sample content should be predicted. If the sample concentration is too high, the sample should be diluted to ensure that the diluted sample meets the detection range of the reagent kit. When calculating, it should be multiplied by the corresponding dilution factor.

1. Sample addition: Set up blank holes, standard holes, and sample holes for testing. Add 100 μ l of sample diluent to the blank well, and add 100 μ l of standard or test sample to the remaining wells respectively. Be careful not to have bubbles. Add the sample to the bottom of the enzyme-linked immunosorbent assay (ELISA) plate well, try not to touch the well wall, gently shake and mix. Cover the ELISA plate with a lid or film, and react at 37 ℃ for 120 minutes. To ensure the validity of the experimental results, please use a new standard solution for each experiment.

2. Discard the liquid, shake dry, do not wash. Add 100 μ l of detection solution A working solution to each well (prepared within one hour before use), cover the enzyme-linked immunosorbent assay plate with a film, and react at 37 ℃ for 60 minutes.

3. After incubating for 60 minutes, discard the liquid in the well, shake dry, wash the plate 3 times, soak for 1-2 minutes each time, approximately 400 μ l/well, shake dry (or tap lightly to dry the liquid in the well).

4. Add 100 μ l of detection solution B working solution (the same as detection solution A working solution) to each well, and react the enzyme-linked immunosorbent assay (ELISA) plate with a film at 37 ℃ for 60 minutes.

5. After incubating for 60 minutes, discard the liquid in the well, shake dry, wash the plate 5 times, soak for 1-2 minutes each time, 350 μ l/well, shake dry (or tap lightly to dry the liquid in the well).

6. Add 90 μ l of substrate solution to each well in sequence, and coat the enzyme-linked immunosorbent assay (ELISA) plate with a film at 37 ℃ to avoid light for color development (within 30 minutes, if there is a clear gradient blue color visible to the naked eye in the first 3-4 wells of the standard sample, and the gradient is not obvious in the last 3-4 wells, it can be terminated).

7. Add 50 μ l of termination solution to each well in sequence to terminate the reaction, at which point the blue color will turn yellow. The order of adding termination solution should be as similar as possible to the order of adding substrate solution. To ensure the accuracy of the experimental results, the termination solution should be added as soon as the substrate reaction time is up.

8. Use an enzyme-linked immunosorbent assay (ELISA) to measure the optical density (OD) of each well in sequence at a wavelength of 450nm. Immediately conduct testing after adding termination solution.

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