ACE2, Human, BioAssay(TM) ELISA Kit (Angiotensin-converting Enzyme 2, ACE-related Carboxypeptidase, Angiotensin-converting Enzyme Homolog, ACEH, Metalloprotease MPROT15, Processed Angiotensin-converting Enzyme 2, UNQ868/PRO1885)

By EST database searching for sequences showing homology to the zinc metalloprotease angiotensin-I converting enzyme, a full-length ACE2 cDNA, originally called ACEH, was isolated, which encoded a deduced 805aa protein that shares approximately 40% identity with the N- and C-terminal domains of ACE. ACE2 contains a potential 17aa N-terminal signal peptide and a putative 22aa C-terminal membrane anchor. Northern blot analysis detected high expression of ACE2 in kidney, testis, and heart, and moderate expression in colon, small intestine, and ovary. The soluble, truncated form of ACE2 lacks the transmembrane and cytosolic domains in CHO cells and is glycosylated protein that was able to cleave angiotensin I and angiotensin II , but not bradykinin. ACE converts angiotensin I to angiotensin II, which has 8aa, ACE2 converts angiotensin I to angiotensin 1-9, which has 9aa. This can then further be converted by ACE to a shorter peptide, angiotensin 1-7, which is a blood vessel dilator. Using ACE2 null mice, Crackower et al. showed that ACE2 was critically involved in a cardiac contractility. This assay is a sandwich ELISA for quantitative determination of human ACE2 in biological fluids. A polyclonal antibody specific for ACE2 has been precoated onto the 96-well microtiter plate. Standards and samples are pipetted into the wells for binding to the coated antibody. After extensive washing to remove unbound compounds, ACE2 is recognized by the addition of a biotinylated polyclonal antibody specific for ACE2 (Detection Antibody). After removal of excess biotinylated antibody, HRP labeled streptavidin (Detector) is added. Following a final washing, peroxidase activity is quantified using the substrate 3,3',5,5'-tetramethylbenzidine (TMB). The intensity of the color reaction is measured at 450nm after acidification and is directly proportional to the concentration of ACE2 in the samples. The assay range is 0.391-25ng/ml ACE2/ml. The lowest level of ACE2 that can be detected by this assay is 293pg/ml.\n\nSample Type:\nCell and tissue culture supernatants, urine, plasma, serum, as well as many other biological fluids.\n\nIntended Use:\nFor the quantitative determination of human ACE2 in biological fluids.\n\nSensitivity:\n293pg/ml\n\nRange:\n0.391-25ng/ml ACE2/ml\n\nSpecificity:\nMammalian\n\nTest Principle:\nThis assay is a sandwich ELISA for quantitative determination of human ACE2 in biological fluids. A polyclonal antibody specific for ACE2 has been precoated onto the 96-well microtiter plate. Standards and samples are pipetted into the wells for binding to the coated antibody. After extensive washing to remove unbound compounds, ACE2 is recognized by the addition of a biotinylated polyclonal antibody specific for ACE2 (Detection Antibody). After removal of excess biotinylated antibody, HRP labeled streptavidin (Detector) is added. Following a final washing, peroxidase activity is quantified using the substrate 3,3',5,5'-tetramethylbenzidine (TMB). The intensity of the color reaction is measured at 450nm after acidification and is directly proportional to the concentration of ACE2 in the samples.\n\nKit Components:\nPre-coated Microtiter Plate: 12x8 well strips\nWash Buffer (10X): 1x50ml\nDiluent (5X):1x50ml\nDetection Antibody: 1x12ml\nDetector 100X (HRP Labeled Streptavidin): 1x150ul\nHuman ACE2 Standard (lyophilized): 50ng\nHuman ACE2 QC Sample (lyophilized): 1 vial\nSubstrate Solution (TMB): 1x12ml\nStop Solution: 1x12ml\nPlate Sealers (plastic film): 3 each\n\nStorage and Stability:\nStore powder at 4 degrees C liquid at -20 degrees C. Store other components at 4 degrees C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

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