Description
Background: Angiotensin-converting enzyme (ACE) is a zinc-containing dipeptidyl carboxypeptidase widely distributed in mammalian tissues and is thought to play a critical role in blood pressure regulation. The predicted protein is identical, from residue 37 to its C terminus, to the second half or C-terminal domain of the endothelial ACE sequence. The protein sequence inferred consists of a 732-residue preprotein including a 31-residue signal peptide. The mature polypeptide has a molecular weight of 80,073. Although ACE has been studied primarily in the context of its role in blood pressure regulation, this widely distributed enzyme has many other physiological functions. The ACE gene encodes two isozymes. The somatic isozyme is expressed in many tissues, including vascular endothelial cells, renal epithelial cells, and testicular Leydig cells, whereas the testicular or germinal angiotensin-converting enzyme is expressed only in sperm. The standard product used in this kit is recombinant human ACE, consisting of 30-1261 amino acids with the molecular mass of 120KDa.
Principle of the Assay: The Picokine™ Human ACE Pre-Coated ELISA (Enzyme-Linked Immunosorbent Assay) kit is a olid-phase immunoassay specially designed to measure Human ACE with a 96-well strip plate that is pre-coated with antibody specific for ACE. The detection antibody is a biotinylated antibody specific for ACE. The capture antibody is monoclonal antibody from rat and the detection antibody is polyclonal antibody from goat. The kit contains recombinant Human ACE with immunogen: Expression system for standard: NS0; immunogen sequence: L30-L1261. The kit is analytically validated with ready-to-use-reagents. To measure Human ACE, add standards and samples to the wells, then add the biotinylated detection antibody. Wash the wells with PBS or TBS buffer, and add Avidin-Biotin-Peroxidase Complex (ABC-HRP). Wash away the unbounded ABC-HRP with PBS or TBS buffer and add TMB. TMB is an HRP substrate and will be catalyzed to produce a blue color product, which changes into yellow after adding the acidic stop solution. The absorbance of the yellow product at 450nm is linearly proportional to Human ACE in the sample. Read the absorbance of the yellow product in each well using a plate reader, and benchmark the sample wells' readings against the standard curve to determine the concentration of Human ACE in the sample