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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 body fluids.
Principle of the Assay: The MCP-1 concentration in the sample is determined using a sandwich enzyme-linked immunosorbent assay (ELISA) method. A microtiter plate is pre-coated with a specific monoclonal antibody against rat MCP-1, which acts as the solid-phase capture antibody. After incubation with the sample, MCP-1 present in the sample binds to the immobilized antibody. A detection antibody labeled with horseradish peroxidase (HRP) is then added, forming a complex of antibody-antigen-enzyme-labeled antibody. Following a washing step to remove unbound components, the substrate TMB (3,3',5,5'-tetramethylbenzidine) is introduced. Under the catalytic action of HRP, TMB is oxidized to a blue product, which changes to yellow upon the addition of an acidic stop solution. The intensity of the color developed is directly proportional to the amount of MCP-1 in the sample. The optical density (OD) is measured at 450 nm using a microplate reader. By comparing the OD values of the test samples with those of a standard curve, the concentration of MCP-1 in the original sample can be accurately calculated. This method offers high specificity, sensitivity, and reproducibility, making it suitable for both research and diagnostic applications.