E el h0101c

The content of Interleukin-4 (IL-4) (E-EL-H0101c, Elabscience) and Interferon γ (IFN-γ) (E-EL-H0108c, Elabscience) was detected by Enzyme linked immunosorbent assay (ELISA). Briefly, the ELISA kit was taken out and then kept for 15 minutes. Then samples and standards were supplemented to corresponding wells and cultured for 3 hours. Later, the liquid in the ELISA plate was discarded, and eluent was added for washing the plate three times. Biotin-labeled working solution was added for incubation for 1 hour. Then the liquid in the plate was discarded, and the eluent was added to wash for three times. The enzyme binding working solution was added and cultured for half an hour, and 100 μl of color developing solution was supplemented for incubation in dark at room temperature for 15 minutes. 100 μl of stop solution was added and mixed. The absorbance value was detected within 30 minutes. A standard curve was drawn, and the sample concentration was calculated.

The expression of COX-2 (E-EL-H1846c, Elabscience), IL-4 (E-EL-H0101c, Elabscience), and IL-17a (EK117/2-02, MULTISCIENCES) levels in patients’ serum was detected using ELISA. All the serum samples were diluted 10 times. Then 100 μl each dilution of standard, blank, and sample was added into 96-well plates. After adding biotinylated detection Ab working solution, the wash buffer was used for washing. Moreover, HRP conjugate working solution and substrate reagent were then added for reaction. The stop solution was finally applied for assessment.

The levels of IL-4 (Elabscience, #E-EL-H0101c), IL-5 (Elabscience, #E-EL-H0191c), IL-13 (Elabscience, #E-EL-H0104c), and TSLP (Elabscience, #E-EL-H1598c; Jonln, #JL20618-96T) in the cell supernatants were measured by the ELISA kit following the manufacturer’s instructions. Concentration was calculated according to the standard curve.