ELISA kit

ELISA kitPrecautions

1. Strictly follow the prescribed time and temperature for incubation to ensure accurate results. All reagents must reach room temperature before use20-25℃Immediately refrigerate and store the reagents after use.

2. 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 on the bottom of the board, otherwise it will affectODValue.

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 reagent 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.

20Dilution of washing buffer: distilled water1：20Dilution, i.e1copy20×Add washing buffer solution19Distilled water.

ELISA kitoperating steps

1. Balance from room temperature20minTake out the required Flat noodles from the rear aluminum foil bag, and seal the remaining Flat noodles with a self sealing bag4℃.

2. Set up standard wells and sample wells, and add different concentrations of standard samples to each standard well50μL；

3. Sample wellmiddleplusentertest sample50μLBlank holes are not added.

4. Add horseradish peroxidase to each well of the standard and sample wells, except for blank wells（HRP）Labeled detection antibodies100μLSeal the reaction hole with a sealing film,37℃Incubate in a water bath or constant temperature box60min.

5. Discard the liquid, pat dry on absorbent paper, and fill each hole with detergent（350μL）Let it stand still1minDiscard the detergent, pat dry on absorbent paper, and repeat the washing process5The board can also be washed using a board washing machine.

6. Add substrate to each wellATheBeach50μL，37℃Incubate in the dark15min.

7. Add termination solution to each well50μL，15minInside, within450nmMeasure the wavelength of each holeODValue.

Calculation of experimental results

withThe measured standardODvalueFor the horizontal axis,Concentration of standard samplesThe value is on the vertical axis, on the coordinate paperOr use relevant software to drawstandard curveAnd obtainLinear regression equation，Take the sampleODSubstitute the value into the equation and calculate the sampleofconcentration.