Mouse tnf alpha elisa max standard

The inflammation-related cytokines were determined from plasma using the Mouse IL-6 ELISA MAX™ Standard, Mouse TNF-alpha ELISA MAX Standard, and Mouse MCP-1 ELISA MAX Standard (Biolegend, San Diego, CA, USA) according to the manufacturer's instructions. In brief, samples and standards were added to a pre-coated 96-well plate and incubated for 2 h. Next, either anti-mouse IL-6, TNFα, or MCP1 antibody was added, and the mixture was incubated for 2 h. Detection antibody was then added to each well, and the mixture was incubated at RT for 1 h. Following incubation with Avidin-HRP solution at RT for 30 minutes, TMB Substrate Solution was added, and the mixture was incubated at RT for 20 min. The reactions were terminated using Stop Solution, and absorbance was read at 450 nm using an ELISA reader (BioTek, Winooski, VT, USA).