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EK000081-EK0557: Human ACE ELISA Kit
Range: 156pg/ml-10,000pg/ml
Sensitivity < 5 pg/ml
Specificity: no detectable cross-reactivity with any other cytokine.
Application: for quantitive detection of human ACE in sera, plasma, body fluids, tissue lysates or cell culture supernates.
Expiration: Four months at 4°C and eight months at -20°C.
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 80,073.1 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.2 The standard product used in this kit is recombinant human ACE, consisting of 30-1261 amino acids with the molecular mass of 120KDa.
Principle
The ELISA Kit is based on standard sandwich enzyme-linked immune-sorbent assay technology. The target specific antibodies are precoated onto 96-well plates. The target from the sample is bound to the microwell. The biotinylated target specific detection antibodies are added to the microwells and followed by washing with the PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex is added and unbound conjugates are washed away with the PBS or TBS buffer. TMB, the HRP (horseradish peroxidase) substrate, is used to visualize color change resulting from the HRP enzymatic reaction. TMB is catalyzed by HRP to produce the blue color. The color changes into yellow after the acidic stop solution is added. The density of the yellow color is proportional to the target amount from the sample captured in the microwells.
Reference
1. Ehlers, M. R. W.; Fox, E. A.; Strydom, D. J.; Riordan, J. F. Molecular cloning of human testicular angiotensin-converting enzyme: the testis isozyme is identical to the C-terminal half of endothelial angiotensin-converting enzyme. Proc. Nat. Acad. Sci. 86: 7741-7745, 1989.
2. Ramaraj, P.; Kessler, S. P.; Colmenares, C.; Sen, G. C. Selective restoration of male fertility in mice lacking angiotensin-converting enzymes by sperm-specific expression of the testicular isozyme. J. Clin. Invest. 102: 371-378, 1998.
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