Intracellular Staining by Flow Cytometry
|Detection of ECE‑1 in MCF‑7 Human Cell Line by Flow Cytometry. MCF‑7 human breast cancer cell line was stained with Rat Anti-Human ECE‑1 Monoclonal Antibody (Catalog # MAB17841, filled histogram) or isotype control antibody (Catalog # MAB005, open histogram), followed by Allophycocyanin-conjugated Anti-Rat IgG F(ab')2 Secondary Antibody (Catalog # F0113). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.|
Endothelin-converting Enzyme 1 (ECE-1) is a zinc protease of the neprilysin (NEP) family, which also includes ECE-2, PEX, XCE, DINE, Kell and several NEP-like proteins (1). ECE-1 is a type II transmembrane protein with a short cytoplasmic tail and a large ectodomain. Four alternatively spliced isoforms differ in their cytoplasmic tail (2, 3). In addition to big endothelin-1, ECE-1 cleaves a variety of bioactive peptides such as bradykinin, neurotensin, angiotensin I, and substance P (1). Together with ECE-2, it is also involved in degradation of beta -amyloid peptide (4). The ectodomain of human ECE-1, which is common to all isoforms, was expressed with an N-terminal His tag and purified.
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