Human IL-10 ELISA Kit

From Assay Genie, this Sandwich ELISA Kit (Double Antibody) is designed for the quantitative measurement of human IL-10 (synonyms: IL-10, IL10, IL10A, CSIF, TGIF). It uses a colorimetric detection method and is suitable for serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid samples. The assay has a sensitivity of 4.688 pg/mL and a standard curve range of 7.813–500 pg/mL (1 ng = 14500 mIU).

Procedure: equilibrate reagents and TMB substrate to room temperature, then load standards, samples and blank/zero controls onto the pre-coated plate and incubate at 37 °C for 90 minutes; add biotinylated detection antibody and incubate at 37 °C for 60 minutes; add HRP-streptavidin (SABC) and incubate at 37 °C for 30 minutes; add TMB substrate and incubate in the dark for 10–20 minutes; add stop solution and read absorbance at 450 nm. Sample-to-sample linearity at 1:2, 1:4 and 1:8 dilutions was 86–101%, 96–100% and 81–95% for serum (n = 5); 88–102%, 93–100% and 81–96% for EDTA plasma (n = 5); and 92–102%, 90–98% and 80–95% for heparin plasma (n = 5). Spike-and-recovery averages were 92% for serum (range 86–97%, n = 5), 100% for EDTA plasma (range 93–105%, n = 5) and 96% for heparin plasma (range 90–102%, n = 5).

Sample preparation: for serum, allow blood to clot and centrifuge at 1000 × g for 20 minutes; for plasma, collect in anticoagulant tubes and centrifuge at 1000 × g for 15 minutes at 2–8 °C; for tissue, homogenize in PBS with protease inhibitors; for cell culture supernatant, centrifuge at 2500 rpm for 5 minutes; for cell lysates, lyse cells using lysis buffer with protease inhibitors. Unopened kit components should be stored at 2–8 °C for up to 12 months.