Mouse
EpCAM/TROP-1 Antibody Summary
Gln24-Thr266
Accession # Q99JW5
Applications
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Scientific Data
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Detection of EPCAM in Mouse Colon via seqIF™ staining on COMET™ EPCAM was detected in immersion fixed paraffin-embedded sections of Mouse Colon using Rabbit Anti-Mouse EPCAM, Monoclonal Antibody (Catalog # MAB11679) at 0.5 μg/mL at 37 ° Celsius for 4 minutes. Before incubation with the primary antibody, tissue underwent an all-in-one dewaxing and antigen retrieval preprocessing using PreTreatment Module (PT Module) and Dewax and HIER Buffer H (pH 9; Epredia Catalog # TA-999-DHBH). Tissue was stained using the Alexa Fluor™ Plus 647 Goat anti-Rabbit IgG Secondary Antibody at 1:200 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR647RB) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the membrane. Protocol available in COMET™ Panel Builder.
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Detection of Mouse EpCAM/TROP‑1 by Western Blot. Western Blot shows lysates of mouse colon tissue. PVDF membrane was probed with 1 µg/ml of Rabbit Anti-Human EpCAM/TROP‑1 Monoclonal Antibody (Catalog # MAB11679) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for EpCAM/TROP‑1 at approximately 40 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.
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Detection of
EpCAM/TROP‑1 in Mouse Colon.
EpCAM/TROP‑1 was detected in immersion fixed paraffin-embedded sections of mouse colon using Rabbit Anti-Human
EpCAM/TROP‑1 Monoclonal Antibody (Catalog # MAB11679) at 3 µg/ml for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003) or the HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to the membrane. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
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Detection of
EpCAM/TROP‑1 in Mouse Lung.
EpCAM/TROP‑1 was detected in immersion fixed paraffin-embedded sections of mouse lung using Rabbit Anti-Human
EpCAM/TROP‑1 Monoclonal Antibody (Catalog # MAB11679) at 3 µg/ml for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003) or the HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to the membrane. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
Preparation and Storage
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: EpCAM/TROP1
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 squamousal 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).
- Balzar, M. et al. (1999) J. Mol. Med. 77:699.
- Boer, C.J. et al. (1999) J. Pathol. 188:201.
- Litvinow, S.V. et al. (1994) J. Cell Biol. 125:437.
- Balzar, M. et al. (2001) Mol. Cell. Biol. 21:2570.
- Balzar, M. et al. (1998) Mol. Cell. Biol. 18:4388.
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