ANGPTL3, Mouse, BioAssay(TM) ELISA Kit (Angiopoietin-related Protein 3, Angiopoietin-like Protein 3)

The angiopoietins are a family of growth factors that are specific for vascular endothelium. The full-length cDNA encoding angiopoietin-like protein 3 (ANGPTL3) from a human fetal liver/spleen cDNA library has 460aa and the characteristic structure of angiopoietins: a signal peptide, an extended helical domain predicted to form dimeric or trimeric coiled-coils, a short linker peptide, and a globular fibrinogen-like domain (FLD). Human ANGPTL3 shares 76% aa sequence identity with mouse Angptl3. Northern blot analysis of human tissues showed a preferential expression of 4 ANGPTL3 transcripts being 4.5, 3.0, 2.8, and 1.7kb in liver. ANGPTL3 can induce angiogenesis in the rat corneal assay. The FLD alone was sufficient to induce endothelial cell adhesion and in vivo angiogenesis. Microarray analysis showed that mouse hematopoietic stem cell (HSC)-supportive fetal liver CD3-positive cells expressed Angptl2 and Angptl3. The ANGPTL3 ELISA Kit is to be used for the in vitro quantitative determination of mouse/ rat ANGPTL3 in biological fluids. This assay is a sandwich ELISA for quantitative determination of mouse or rat ANGPTL3 in biological fluids. A polyclonal antibody specific for ANGPTL3 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, ANGPTL3 is recognized by the addition of a biotinylated polyclonal antibody specific for ANGPTL3 (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 ANGPTL3 in the samples. The assay range is 0.016-1ng/ml ANGPTL3/ml. The lowest level of ANGPTL3 that can be detected by this assay is 15pg/ml.\n\nSample Type:\nCell and tissue culture supernatants, urine, plasma, serum, as well as many other biological fluids.\n\nIntended Use:\nFor the in vitro quantitative determination of mouse/ rat ANGPTL3 in biological fluids.\n\nSensitivity:\n15pg/ml \n\nRange:\n0.016-1ng/ml ANGPTL3/ml\n\nSpecificity:\nMammalian\n\nTest Principle:\nThis assay is a sandwich ELISA for quantitative determination of mouse or rat ANGPTL3 in biological fluids. A polyclonal antibody specific for ANGPTL3 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, ANGPTL3 is recognized by the addition of a biotinylated polyclonal antibody specific for ANGPTL3 (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 ANGPTL3 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\nMouse ANGPTL3 Standard (lyophilized): 1 vial\nMouse ANGPTL3 QC Sample (lyophilized): 1 vial\nSubstrate Solution (TMB): 1x6ml\nStop Solution: 1x6ml\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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