Mouse Myoglobin (MYO/MB) Kit ELISA Procedure:

(1) Add 100 μ l of the test sample to each well of the test sample, and set 3 parallel wells for each type of sample; Set up two negative control wells, add untreated samples to each well

Cell lysate of group 100 μ 1; Set up another blank control well and add 100 μ 1 of pure cell lysate.

(2) Place the enzyme-linked immunosorbent assay (ELISA) plate at 4 ℃ and coat overnight.

(3) Plate washing: Absorb the reaction solution in the hole, rinse it once with washing solution (fill the plate hole with washing solution, then shake it off), and then fill the plate with washing solution

Soak the hole for 1-2 minutes and shake intermittently. Shake off the liquid inside the hole and pat dry on absorbent paper. Repeat washing 3-4 times.

(4) Add PBS 50 μ 1 to each negative control well, and add 50 μ 1 of 1:500 diluted anti human AIF antibody working solution to each sample well and blank well.

(5) Place the enzyme-linked immunosorbent assay (ELISA) plate in a wet box at 37 ℃ and incubate for 60 minutes.

(6) Wash the board, same as (4).

(7) Add 100 μ l of HRP labeled goat anti immune antibody working solution diluted 1:5000 to each well.

(8) Place the enzyme-linked immunosorbent assay (ELISA) plate in a wet box at 37 ℃ and incubate for 60 minutes.

(9) Wash the board, same as (4).

(10) Add 100 μ l of TMB chromogenic solution to each well, gently mix for 10 seconds, and let it react in the dark at 37 ℃ for 15-20 minutes.

(11) Add 100 μ 12mo1/LH2S04 to each well to terminate the reaction.

(12) Measure the absorbance values V1 and V2 at 450nm and 630nm respectively, and the final measured 0D value is the difference between the two (V1-v2) to reduce the impact on the container

Light interference caused by scratches or fingerprints.

(13) Data processing: After obtaining the OD values of the specimen (S) and negative control (N), calculate the S/N value. S/N>2.1 is the positive judgment criterion.