Rat C5a (Complement Component 5a) Microsample ELISA Kit

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Rat C5a. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Rat C5a. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Rat C5a, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Rat C5a in the samples is then determined by comparing the OD of the samples to the standard curve.Detection Range: 1.57-100 ng/mL. Sensitivity: 0.63 ng/mL. Assay Type: Sandwich. Standard: 100 ng/mL. Sample Type: sSerum, plasma and other biological fluids. Assay duration: 3.5h. Protein Function: Precursor of the C5a anaphylatoxin and complement C5b components of the complement pathways, which consist in a cascade of proteins that leads to phagocytosis and breakdown of pathogens and signaling that strengthens the adaptive immune system. Activated downstream of classical, alternative, lectin and GZMK complement pathways , Component of the membrane attack complex (MAC), a multiprotein complex activated by the complement cascade, which inserts into a target cell membrane and forms a pore, leading to target cell membrane rupture and cell lysis. Complement C5b is generated following cleavage by C5 convertase and initiates formation of the MAC complex: C5b binds sequentially C6, C7, C8 and multiple copies of the pore-forming subunit C9. During MAC complex assembly, the C5b6 subcomplex, composed of complement C5b and C6, associates with the outer leaflet of target cell membrane, reducing the energy for membrane bending , Mediator of local inflammatory process released following cleavage by C5 convertase. Acts by binding to its receptor (C5AR1 or C5AR2), activating G protein-coupled receptor signaling and inducing a variety of responses including intracellular calcium release, contraction of smooth muscle, increased vascular permeability, and histamine release from mast cells and basophilic leukocytes. C5a is also a potent chemokine which stimulates the locomotion of polymorphonuclear leukocytes and directs their migration toward sites of inflammation [The Uniprot Consortium]