Enzyme Activity of Plant Cysteine Protease 3 (Casp-3) ELISA Kit

Enzyme Activity of Plant Cysteine Protease 3 (Casp-3) ELISA KitPrecautions

1. Strictly follow the prescribed time and temperature for incubation to ensure accurate results. All reagents must reach room temperature of 20-25 ℃ before use. Immediately refrigerate and store the reagents after use.

Incorrect board washing can lead to inaccurate results. Ensure to absorb as much liquid as possible from the well before adding the substrate. Do not let the micropores dry out during the incubation process.

3. Eliminate residual liquid and fingerprints at the bottom of the board, otherwise it will affect the OD value.

4. The substrate color solution should be colorless or very light in color, and the substrate solution that has turned blue cannot be used.

5. Avoid cross contamination between reagents and specimens to prevent erroneous results.

6. Avoid direct exposure to strong light during storage and incubation.

7. After balancing to room temperature, open the sealing bag to prevent water droplets from condensing on the cold Flat noodles.

8. Any reaction reagents should not come into contact with bleach solvents or the strong gases emitted by bleach solvents. Any bleaching component will destroy the biological activity of the reactants in the reagent kit.

9. Expired products cannot be used.

10. If there is a possibility of spreading the disease, all samples should be managed and processed according to the prescribed procedures and testing devices.

Reagent preparation

The reagent kit should be taken out of the refrigerated environment and equilibrated at room temperature before use.

Dilution of 20 x washing buffer: Dilute distilled water at a ratio of 1:20, that is, 1 part of 20 x washing buffer is added to 19 parts of distilled water.

Enzyme Activity of Plant Cysteine Protease 3 (Casp-3) ELISA Kitoperating steps

1. Take out the required Flat noodles from the aluminum foil bag after 20 min of room temperature balance, and seal the remaining Flat noodles with a self sealing bag and put it back at 4 ℃.

2. Set up standard wells and sample wells, and add 50 μ L of standard samples of different concentrations to each standard well;

3. Add 50 μ L of the sample to be tested into the sample well; Blank holes are not added.

4. Except for blank wells, add 100 μ L of horseradish peroxidase (HRP) labeled detection antibody to each well of the standard and sample wells, seal the reaction well with a sealing plate membrane, and incubate at 37 ℃ in a water bath or constant temperature box for 60 minutes.

5. Discard the liquid, pat dry on absorbent paper, fill each well with washing solution (350 μ L), let it stand for 1 minute, shake off the washing solution, pat dry on absorbent paper, repeat washing the board 5 times (or use a washing machine to wash the board).

6. Add 50 μ L of substrate A and B to each well, and incubate at 37 ℃ in the dark for 15 minutes.

7. Add 50 μ L of termination solution to each well and measure the OD value of each well at a wavelength of 450nm within 15 minutes.

Calculation of experimental results

Using the OD value of the measured standard as the horizontal axis and the concentration value of the standard as the vertical axis, draw a standard curve on a coordinate paper or with relevant software, and obtain a linear regression equation. Substitute the OD value of the sample into the equation to calculate the concentration of the sample.