Human EpCAM/TROP-1 Antibody AF960: R&D Systems

Human
EpCAM/TROP-1 Antibody

(4 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human EpCAM/TROP-1 in direct ELISAs and Western blots. In direct ELISAs, less than 1% cross-reactivity with recombinant human TROP-2 is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human EpCAM/TROP-1, extracellular domain
    Gln24-Lys265
    Accession # CAA32870
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.2 µg/mL
    See below
  • Flow Cytometry
    0.25 µg/106 cells
    See below
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Immunocytochemistry
    5-15 µg/mL
    See below
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.
Data Examples
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).
Immunocytochemistry
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.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 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 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).

  • References:
    1. Balzar, M. et al. (1999) J. Mol. Med. 77:699.
    2. Boer, C.J. et al. (1999) J. Pathol. 188:201.
    3. Litvinow, S.V. et al. (1994) J. Cell Biol. 125:437.
    4. Balzar, M. et al. (2001) Mol. Cell. Biol. 21:2570.
    5. Balzar, M. et al. (1998) Mol. Cell. Biol. 18:4388.
  • Long Name:
    Epithelial Cell Adhesion Molecule
  • Entrez Gene IDs:
    4072 (Human); 17075 (Mouse)
  • Alternate Names:
    17-1A; 323/A3; ACSTD1; antigen identified by monoclonal AUA1; CD326 antigen; CD326; Cell surface glycoprotein Trop-1; chromosome 4, surface marker (35kD glycoprotein); DIAR5; EGP; EGP-2; EGP314; EGP40; EpCAM; epithelial cell adhesion molecule; Epithelial cell surface antigen; Epithelial glycoprotein 314; Epithelial glycoprotein; ESA; GA733-2; GA733-2EGP34; gp40; hEGP314; HNPCC8; KS 1/4 antigen; KS1/4; KSAHEA125; M1S2; M4S1; M4S1Ly74; Major gastrointestinal tumor-associated protein GA733-2; MIC18MH99; MOC31; TACST-1; TACSTD1; TROP1; TROP1CD326; Tumor-associated calcium signal transducer 1CO-17A
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

4 Citations: Showing 1 - 4
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Species
Applications
Sample Type
  1. Fast and selective cell isolation from blood sample by microfiber fabric system with vacuum aspiration
    Sci Technol Adv Mater, 2016;17(1):807-815.
    Species: Human
    Sample Type: Whole Cells
    Application: Cell selection
  2. HER2 and EGFR overexpression support metastatic progression of prostate cancer to bone
    Cancer Res., 2016;0(0):.
    Species: Human
    Sample Type: Whole Blood
    Application: CTC
  3. EpCAM-Regulated Transcription Exerts Influences on Nanomechanical Properties of Endometrial Cancer Cells that Promote Epithelial-to-Mesenchymal Transition
    Cancer Res, 2016;0(0):.
    Species: Human
    Sample Type: Whole Cells
    Application: Cell Surface Adhesion
  4. Stromal gene expression defines poor-prognosis subtypes in colorectal cancer.
    Authors: Calon A, Lonardo E, Berenguer-Llergo A, Espinet E, Hernando-Momblona X, Iglesias M, Sevillano M, Palomo-Ponce S, Tauriello D, Byrom D, Cortina C, Morral C, Barcelo C, Tosi S, Riera A, Attolini C, Rossell D, Sancho E, Batlle E
    Nat Genet, 2015;47(4):320-9.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC
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