Human TACE/ADAM17 Ectodomain Antibody

Detection of Human TACE/ADAM17 by Flow Cytometry

Effects of the engineered S197P mutation on CD16a shedding in NK cells.NK92 cells transduced with empty vector (vector only), CD16a, or CD16a/S197P were treated without (Unstim.) or with PMA (100ng/ml) for 30 minutes at 37°C (A), with IL-12 and IL-18 (100ng/ml and 400ng/ml, respectively) for 24 hours at 37°C (B), or with Raji cells and rituximab for 60 min at 37°C (C). Cell surface levels of CD16a were determined by flow cytometry. Isotype-matched negative control antibody staining is indicated by a dotted line. (D) Parent NK92 cells and transduced cells expressing CD16a or CD16a/S197P were treated with Raji cells and rituximab in the presence or absence of the ADAM17 inhibitor BMS566394 (5μM) for 60 min at 37°C. Soluble CD16a levels were determined by ELISA. Each treatment condition was repeated 3 times and the data are representative of 3 independent experiments. Bar graphs show mean ± SD. Statistical significance is indicated as ***P<0.001. (E) NK92 cells expressing CD16a or CD16a/S197P were stained with the anti-ADAM17 mAbs M220, 623, 633, or an isotype-matched negative control antibody, as indicated. (F) CD56+CD45+ NK cells derived from mock-transduced iPSCs (left panel) or iPSCs expressing recombinant CD16a or CD16a/S197P (right panels) were incubated with or without K562 target cells for 4 hours at 37°C. For all histogram plots, the x-axis = Log 10 fluorescence, the y-axis = cell number, and the data are representative of at least 3 independent experiments. Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0121788), licensed under a CC-BY license. Not internally tested by R&D Systems.