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Monocyte chemoattractant protein-1 (MCP-1) Assay Principle
Objective: This kit is designed to quantify the levels of monocyte chemoattractant protein-1 (MCP-1) in various biological samples, including serum, plasma, tissue homogenates, and other related fluids.
Principle of the Assay: The MCP-1 concentration is determined using a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA). The process begins by coating microtiter plate wells with a specific monoclonal antibody against rat MCP-1. After incubation, the sample containing MCP-1 is added to the wells, allowing the antigen to bind to the immobilized antibody. A horseradish peroxidase (HRP)-labeled secondary antibody is then introduced, which binds to the captured MCP-1, forming a stable immune complex. Following a thorough wash step to remove unbound components, a TMB (3,3',5,5'-tetramethylbenzidine) substrate is added. Under the catalytic action of HRP, TMB turns blue, and the reaction is stopped by adding an acidic solution, resulting in a yellow color. The intensity of the color is directly proportional to the amount of MCP-1 present in the sample. The optical density (OD) is measured at 450 nm using a microplate reader, and the concentration of MCP-1 is calculated based on a standard curve generated from known concentrations of the protein. This method ensures high specificity, sensitivity, and reproducibility for MCP-1 detection in various experimental settings.