Fig 1: Validation of EV-CATCHER customized antibodies and characterization of placental EVs. (a). Western blots displaying the specificity of the monoclonal placental alkaline phosphatase antibody against the PLAP recombinant protein (Novus, Centennial, CO, USA, #NBP2-52266) used for EV-CATCHER customization versus the alkaline phosphatase, tissue-nonspecific isozyme antibody against the ALPL recombinant protein (Novus, Centennial, CO, USA, #2909-AP-010). (b). EV-CATCHER assay customized with an anti-PLAP antibody conjugated to a uracilated DNA linker that is connected by a biotin to the bottom of a streptavidin coated well. Using uracil DNA glycosylase (UNG), the immobilized EV-anti-PLAP duplex is released by enzymatic digestion of the double-stranded uracilated DNA linker. (c). Representative Transmission electron microscopy (TEM) images of EVs isolated from placenta previa and placenta percreta maternal plasma, scale bar 100 µm. (d). Representative ONi super resolution images of EVs isolated from placenta previa and placenta percreta maternal plasma displaying both high PLAP+ expressing EVs and low PLAP+ expressing EVs, scale bar of 20 µm for large field-of-view images and 200 nm for individual representative EVs.
Fig 2: Tissue-nonspecific alkaline phosphatase contents of the plasma samples were unchanged. A Levels of tissue-nonspecific alkaline phosphatase (TNAP) in plasma from controls (C) and patients with AAA, including low aortic growth (LG) and high aortic growth (HG). B Alkaline phosphatase (ALP) activity measured with a commercial assay kit in plasma from healthy subjects and aneurysm patients. C Correlation between TNAP protein levels and ALP activity, indicating a strong positive relationship. The data are presented as scatter plot (A, B). Statistical analysis was conducted using the Kruskal-Wallis test with Dunn’s post hoc test. No statistically significant differences were observed between the groups assessed. Asterisks denote statistically significant differences: ***P < 0.001
Supplier Page from R&D Systems, a Bio-Techne Brand for Alkaline Phosphatase/ALPL Protein
Available conjugates: Sizes Available: 10 ug