|Detection of Human EpCAM/TROP‑1 by Western Blot. Western blot shows lysates of HCT‑116 human colorectal carcinoma cell line, MCF‑7 human breast cancer cell line, and BG01V human embryonic stem cells. PVDF membrane was probed with 0.2 µg/mL of Goat Anti-Human EpCAM/TROP‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF960) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for EpCAM/TROP‑1 at approximately 40 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
Detection of EpCAM/TROP‑1 in HT‑29 Human Cell Line by Flow Cytometry.|
HT‑29 human colon adenocarcinoma cell line was stained with Goat Anti-Human EpCAM/TROP‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF960, filled histogram) or isotype control antibody (Catalog # AB-108-C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107).
|EpCAM/TROP‑1 in BG01V Human Embryonic Stem Cells. EpCAM/TROP‑1 was detected in immersion fixed BG01V human embryonic stem cells using Goat Anti-Human EpCAM/TROP‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF960) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm and cell membrane. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.|
Epithelial Cellular Adhesion Molecule (EpCAM), also known as KS1/4, gp40, GA733-2, 17-1A, and TROP-1, is a 40 kDa transmembrane glycoprotein composed of a 242 amino acid (aa) extracellular domain with two epidermal-growth-factor-like (EGF-like) repeats within the cysteine-rich N-terminal region, a 23 aa transmembrane domain, and a 26 aa cytoplasmic domain. Human and mouse EpCAM share 82% aa sequence identity. In human, EpCAM also shares 49% aa sequence homology with TROP-2/EGP-1. During embryonic development, EpCAM is detected in fetal lung, kidney, liver, pancreas, skin, and germ cells. In adults, human EpCAM is detected in basolateral cell membranes of all simple, pseudo-stratified, and transitional epithelia, but is not detected in normal squamous stratified epithelia, mesenchymal tissue, muscular tissue, neuro-endocrine tissue, or lymphoid tissue (1). EpCAM expression has been found to increase in actively proliferating epithelia tissues and during adult liver regeneration (1, 2). EpCAM expression is also found to increase in human malignant neoplasias, with most carcinoma expressing EpCAM including those of arising from squamous epithelia (1). EpCAM has been shown function as a homophilic Ca2+ independent adhesion molecule (3). Homophilic adhesion via EpCAM requires the interaction of both EGF-like repeats, with the first EGF-like repeat mediating reciprocal interaction between EpCAM molecules on opposing cells, while the second repeat is involved in lateral interaction of EpCAM. Lateral interaction of EpCAM lead to the formation of dimers and tetramers (4). During homophilic adhesion the cytoplasmic tail of EpCAM interacts with the actin cytoskeleton via a direct association alpha -actinin (5).
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