Human IL-9 ELISA Kit

For each experiment, a standard curve needs to be set for each microplate, and the specific OD value may vary depending on different laboratories, testers, or equipment. The following example data is for reference only. The sample concentration was calculated based on the results of the standard curve. The minimum detectable concentration of IL-9 is less than 20 pg/mL.

Cell Culture Supernatant: Human peripheral blood mononuclear cells (PBMCs) (1.5 x 10⁶ cells/mL) were cultured in complete RPMI 1640 medium containing 10% fetal bovine serum, 2 mM L-glutamine, and antibiotic supplements (penicillin 100 U/mL with streptomycin sulfate 100 μg/mL). PBMC cells were either left unstimulated or stimulated with 10 μg/mL PHA and cultured for 3 to 5 days. Cell supernatants of cultured cells were collected on day 3 and day 5, stored at≤-20°C. Subsequent analysis involved quantitative measurement of IL-9 concentrations in collected culture supernatants is conducted by using standardized immunoassays.

Cell Culture Supernatant: Human peripheral blood mononuclear cells (PBMCs) (1.5 x 10⁶ cells/mL) were cultured in complete RPMI 1640 medium containing 10% fetal bovine serum, 2 mM L-glutamine, and antibiotic supplements (penicillin 100 U/mL with streptomycin sulfate 100 μg/mL). PBMC cells were either left unstimulated or stimulated with 10 μg/mL PHA and cultured for 3 to 5 days. Cell supernatants of cultured cells were collected on day 3 and day 5, stored at≤-20°C. Subsequent analysis involved quantitative measurement of IL-9 concentrations in collected culture supernatants is conducted by using standardized immunoassays.

Ten replicates of each of 3 samples containing different IL-9 concentrations were tested in one assay. Acceptable criteria: CV＜10%.

Three samples containing different concentrations of IL-9 were tested in independent assays. Acceptable criteria: CV<15%.

Recombinant IL-9 was spiked into 3 human serum samples, and then analyzed. The average recovery of IL-9 for serum samples is 109.2%.