Human il 6 elisa kit

Human IL-6 ELISA Kit (Raybiotech) was used according to the manufacturer’s instructions. Briefly, an aliquot (100 µL) of standard or sample was added to each well, and plates were incubated for 2.5 h at room temperature. An aliquot of 100 µL of prepared biotin antibody was added to each well and plates were incubated for 1 h at room temperature. An aliquot of 100 µL of prepared streptavidin solution was added to each well and plates were incubated for 45 min at room temperature. An aliquot of 100 µL of TMB One-Step Substrate Reagent was added to each well and plates were incubated for 30 min at room temperature. Finally, an aliquot of 50 µL of Stop Solution was added to each well and plates were read immediately by measuring absorbance at 450 nm.

The human angiogenesis array (Raybiotech, USA) was used to analyse the soluble mediators according to the manufacturer’s protocol. A human IL-6 ELISA kit (Raybiotech) was used to determine the concentration of human IL-6 in the medium of different treatments according to the manufacturer’s instructions.

The secretion of tumor necrosis factor (TNF)-α (human TNF-α ELISA kit; cat no. PT518), interleukin (IL)-1β (human IL-1β ELISA kit; cat no. PI305), and IL-6 (Human IL-6 ELISA kit; cat no. PI330) in the NP cell culture supernatant was quantitatively assessed using ELISA kits (RayBiotech, GA, USA) according to the manufacturer’s instructions. The OD values at 450 nm were assessed using a Multiskan Spectrum (Molecular Devices, LLC, Shanghai, China) [27 ].