用ELISA试剂盒(ThermoFisher)测定白细胞介素(IL)-

ELISA kit (ThermoFisher) was used to measure the secretion of interleukin (IL)-6 and IL-8). First, the ELISA 96-well plate was coated with 100 μL IL-6 and IL-8 capture antibody, and incubated overnight at 4C. The ELISA plate was washed with phosphate buffered saline (Wash Buffer) containing 0.05% Tween 20. Add 200μL ELISA/ELISPOT (1×) to each well and incubate for 1h. Then, 100 μL of sample was added to incubate for 2 hours, 100 ml of detection antibody was added to incubate for 1 hour, and 100 μL of streptavidin-horseradish peroxidase (HRP) were incubated for 30 minutes. Finally, add 100μL of tetramethylbenzidine (TMB) solution, incubate for 15min and add 50ul of stop solution to show orange yellow. After the end, use a microplate reader (BioTekEpoch2) to detect the absorbance at a wavelength of 450nm). The concentration of each sample was calculated according to the standard curve of IL-6 and IL-8.

The secretion of lecytolein (IL)-6 and IL-8 was determined using the ELISA kit (ThermoFisher). First, the ELISA96 plate was encum coated with 100 μL IL-6 and IL-8 capture antibodies and incubated overnight at 4C. Wash the ELISA plate with Wash Buffer, which contains 0.05% Twain 20. Add 200 μL ELISA/ELISPOT (1×) to each well and incubate 1h. Then, add 100 sl sample incubation 2h, then add 100 ml detection antibody incubation 1h, 100 streptomycin-spicy root peroxidase (HRP) incubation 30min. Finally, a solution of 100 μL of tetamine (TMB) is added, 15min is incubated and 50ul termination fluid is added to orange. At the end, use a microplate reader (BioTekEpoch2) to detect a light absorption of 450nm. The sample concentrations are calculated according to the standard curves of IL-6 and IL-8.

The secretion of IL-6 and IL-8 was measured by ELISA Kit (Thermo Fisher). ELISA 96 well plates were coated with 100 μ l IL-6 and IL-8 capture antibodies and incubated overnight at 4C. The ELISA plates were washed with 0.05% Tween 20 phosphate buffer. 200 μ l ELISA / ELISPOT (1 ×) was added into each well and incubated for 1 h. Then, 100 μ l samples were added to incubate for 2 h, 100 ml of detection antibody was added for 1 h, and 100 μ l streptavidin horseradish peroxidase (HRP) was incubated for 30 min. Finally, 100 μ l of tetramethylbenzidine (TMB) solution was added and incubated for 15 min, and 50 UL of termination solution was added, showing orange yellow color. After that, the absorbance at 450 nm was detected with a microplate reader (BioTek epoch2). The concentration of each sample was calculated according to the standard curve of IL-6 and IL-8.<br>