Description
Background: C5a is a small peptide molecule in the complement system, formed by the cleavage of complement C5, and has strong chemotactic activity. It is a critical regulatory molecule in inflammation and immune responses, capable of attracting various immune cells (such as neutrophils, monocytes, T cells, and macrophages) to the site of inflammation, and participating in the activation, aggregation, and mediation of immune cells and inflammatory responses. C5a promotes the chemotaxis and activation of immune cells by interacting with its specific receptor C5aR, inducing the activation of intracellular signaling pathways, ultimately resulting in various biological effects such as extracellular matrix degradation, intracellular signal transduction, cell apoptosis, and production of inflammatory mediators. In addition to its role in normal immune responses, excessive production and abnormal activation of C5a are also associated with various diseases such as autoimmune diseases, infectious diseases, cardiovascular diseases, neurological diseases, and cancer.
Principle of the Assay: This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti C5a antibody was pre-coated onto the 96-well plate. The biotin conjugated anti C5a antibody was used as the detection antibody. The standards and pilot samples were added to the wells subsequently. After incubation, unbound conjugates were removed by wash buffer. Then, biotinylated detection antibody was added to bind with C5a conjugated on coated antibody. After washing off unbound conjugates, HRP-Streptavidin was added. After a third washing, TMB substrates were added to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that turned yellow after adding a stop solution. Read the O.D. absorbance at 450nm in a microplate reader. The concentration of C5a in the sample was calculated by drawing a standard curve. The concentration of the target substance is proportional to the OD450 value