C-Reactive Protein (CRP) is a member of the Pentraxin family of proteins that are characterized by a cyclic,non-covalent,pentameric structure. IL-6,IL-1 beta,and glucocorticoids induce hepatic C-Reactive Protein synthesis and release. In humans,C-Reactive Protein is a major acute phase protein,increasing by 1,000-fold within 24 to 48 hours of infection,inflammation or tissue damage. C-Reactive Protein exhibits calcium-dependent binding of its principle ligand,phosphocholine,a constituent of bacterial and fungal cell walls. Upon ligand binding,C-Reactive Protein initiates the activation of the complement cascade and binds Fc gamma RI (CD64) and Fc gamma RIIA (CD32a) on phagocytes to activate phagocytic responses. In mouse,C-Reactive Protein is expressed at very low levels and is not an acute phase reactant.Sample Type:Cell culture supernatants,serum,plasma,and tissueIntended Use:The Human CRP BioAssay ELISA Kit is an enzyme-linked immunosorbent assay for the quantitative detection of Human CRP concentrations in cell culture supernatants,serum,plasma,and tissue.Sensitivity:100pg/mlRange:390pg/ml-25000pg/mlSpecificity:Recognizes human CRP.Sample Volume:100ul/wellTest Principle:This assay employs the Sandwich immunoassay technique. An anti-h CRP monoclonal coating antibody is adsorbed onto microwells. CRP present in the sample or standard binds to antibodies adsorbed to the microwells. Following incubation unbound sample or standard are removed during a wash step. a Biotinylated anti-h CRP antibody is added and binds to CRP captured by the first antibody. Following incubation unbound Biotinylated anti-h CRP antibody is removed during a wash step. A Streptavidin-HRP is added and binds to Biotinylated anti-h CRP antibody. Following incubation unbound Streptavidin-HRP is removed during a wash step. A colored product is formed in proportion to the amount of CRP present in the sample. The reaction is terminated by addition of acid and absorbance is measured at 450nm. A standard curve is prepared from seven CRP standard dilutions and CRP sample concentration determined.Kit Components:Aluminium pouches with a Microwell Plate: 8x6Standard: 1xVialBiotinylated antibody: 1xVialStreptavidin-HRP: 1xVialAssay Buffer (25ml/Vial): 1xVialWash Buffer Concentrate 20x (30ml/Vial): 1xVialTMB Substrate Solution: 1x6mlStop Solution (12ml/Vial): 1xVialPlate Covers- Adhesive strips: 2xstripsInstruction: 1xStorage and Stability:Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product,centrifuge the original vial after thawing and prior to removing the cap.