Elisa detection kits

Protein levels of IL-1β, IL-6, TNF-α and NF-κBp65 in spinal cord tissues and PC 12 cells were detected by ELISA kit purchased from Thermo Scientific (Shanghai, China). Protein levels of iNOS and COX-2 in spinal cord tissues and PC 12 cells were detected using the ELISA Detection Kits purchased from Abcam (Shanghai, China). The OD values of iNOS, COX-2, IL-1β, IL-6 and TNF-α in each sample were detected at 450 nm wavelength. Only the OD value of NF-κBp65 was measured at 405 nm wavelength.
Irisin levels in the spinal cord and serum were measured with ELISA kit (Phoenix Pharmaceuticals, Burlingame, CA, USA). The lowest detectable concentration of irisin was 1.29 ng/ml (Aydin et al., 2017 (link)). Sample absorbance was read at 450 nm wavelength with an ELX 800 ELISA reader.

Blood samples from patients were collected, and serum was isolated for the measurement of TSLP levels. Serum TSLP levels were measured according to ELISA method using a human TSLP ELISA detection kit (DY1398; R&D Systems, Minneapolis, MN, USA). All samples were measured in duplicates, and the mean concentration was calculated. ELISA was also used for the measurement of cytokine levels in the supernatant fluid collected from the cultured peripheral blood mononuclear cells (PBMCs) subjected to stimulation with lipopolysaccharide (2 μL·mL⁻¹ of cell culture fluid). All ELISA detection kits were purchased from Abcam Co. (Cambridge, UK) for the measurement of cytokines secreted from PBMCs isolated from patients, including the human interleukin (IL)‐1β ELISA Kit (ab46608, Abcam.Co), human IL‐6 ELISA Kit (ab46027, Abcam.Co), human matrix metalloproteinase‐9 ELISA Kit (ab100610, Abcam.Co), human tumor necrosis factor (TNF)‐α ELISA Kit (ab181421) and human IL‐10 ELISA Kit (ab100519,Abcam.Co).