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 Human ECE‑1 Antigen Affinity‑purified Polyclonal Antibody (Catalog # AF1784, filled histogram) or control antibody (Catalog # AB-108-C, open histogram), followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0108). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.|
|ECE‑1 in Human Kidney. ECE‑1 was detected in immersion fixed paraffin-embedded sections of human kidney using Goat Anti-Human ECE‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1784) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific labeling was localized to the endothelial cells in glomeruli. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.|
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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