Human EpCAM/TROP1 Antibody

R&D Systems | Catalog # AF960

R&D Systems

Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Mouse

Applications

Validated:

Immunohistochemistry, Western Blot, Flow Cytometry, Immunocytochemistry, Simple Western, CyTOF-ready

Cited:

Immunohistochemistry, Western Blot, Flow Cytometry, Immunocytochemistry, Adhesion Assay, Bioassay, Capture Assay, Cell Selection, CTC, CyTof, ELISA Capture, FACS

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all EpCAM/TROP1 Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant human EpCAM/TROP-1, extracellular domain
Gln24-Lys265
Accession # CAA32870

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 and approximately 26% cross-reactivity with recombinant mouse EpCAM/TROP-1 is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human EpCAM/TROP1 Antibody

Detection of Human EpCAM/TROP-1 antibody by Western Blot.

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 (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 antibody in HT-29 Human Cell Line antibody by Flow Cytometry.

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 (AB-108-C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (F0107).
EpCAM/TROP-1 antibody in BG01V Human Embryonic Stem Cells by Immunocytochemistry (ICC).

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.
EpCAM/TROP-1 antibody in Human Colon by Immunohistochemistry (IHC-P).

EpCAM/TROP‑1 in Human Colon.

EpCAM/TROP-1 was detected in immersion fixed paraffin-embedded sections of human colon using Goat Anti-Human EpCAM/TROP-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF960) at 0.3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm and plasma membrane. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
EpCAM/TROP-1 antibody in Human Colon by Immunohistochemistry (IHC-P).

EpCAM/TROP‑1 in Human Colon.

EpCAM/TROP-1 was detected in immersion fixed paraffin-embedded sections of human colon using Goat Anti-Human EpCAM/TROP-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF960) at 0.3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm and plasma membrane. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Detection of Human EpCAM/TROP‑1 by Simple WesternTM.

Simple Western lane view shows lysates of A549 human lung carcinoma cell line exosomes, HT‑29 human colon adenocarcinoma cell line exosomes, and COLO 205 human colorectal adenocarcinoma cell line whole cell lysate (WCL), loaded at 0.2 mg/mL. A specific band was detected for EpCAM/TROP‑1 at approximately 52 kDa (as indicated) using 10 µg/mL of Goat Anti-Human EpCAM/TROP‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF960). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Detection of EpCAM/TROP‑1 in HT-29 Human Colon Adenocarcinoma Cell Line (Positive) and Jurkat Human Acute T Cell Leukemia Cell Line (Negative) Cells.

EpCAM/TROP‑1 was detected in immersion fixed HT‑29 human colon adenocarcinoma cell line (positive) and Jurkat human acute T cell leukemia cell line (negative) cells using Goat Anti-Human EpCAM/TROP‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF960) at 5 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 cell surface. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Human EpCAM / TROP1 ELISA Standard Curve

Human EpCAM / TROP1 ELISA Standard Curve

Recombinant Human
EpCAM/TROP‑1 Fc Chimera (Catalog # 960-EP) was serially diluted and captured by Mouse Anti-Human
EpCAM/TROP‑1 Monoclonal Antibody (Catalog # MAB9601) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Human
EpCAM/TROP‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF960) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)
Detection of EpCAM/TROP1 by Immunohistochemistry

Detection of EpCAM/TROP1 by Immunohistochemistry

Laser-scanning confocal images of human colon cross sections with the PCP workflow applied. (A) Representative laser-scanning confocal images, stained with EpCAM (cyan), DAPI (gray), and G3BP1 (magenta). Scale bars represent 50 μm as indicated. (B) Illustrative images of the pipeline’s workflow to identify cells within the lamina propria. The IdentifyPrimaryObjects module was initially applied to delineate nuclei (I) and to recognize the intestinal feature within the whole image (II). The MaskObjects module was then implemented to remove the nuclei of cells that were within the specified intestinal region (III). Subsequently, the IdentifySecondaryObjects module was used to define a cell object by a specific set of pixels from the nucleus as starting point (IV). To identify G3BP1 spots, the IdentifyPrimaryObjects module was employed once more on the G3BP1 immunoreactivity channel (V). Image VI illustrates a comprehensive delineation of G3BP1-positive and -negative cells. The combination of all final objects permits the monitoring of individual cells (cyan for G3BP1-positive and magenta for G3BP1-negative), and G3BP1 spots (yellow) within a final image (VII and VIII). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/41354404), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of EpCAM/TROP1 by Immunohistochemistry

Detection of EpCAM/TROP1 by Immunohistochemistry

Cleavage-resistant EpCAM does not prevent deterioration of intestinal epithelia in HAI-2-deficient mice. (A-C) Representative images (from at least five mice per genotype) of H&E staining (A), and EpCAM (B) and claudin-7 (C) immunostaining of small (top panels) and large (bottom panels) intestines from 5 days old control (left), and HAI-2-deficient (Vil-Cre+/0;Spint2fl/−) mice expressing wildtype (Epcam+/+), R80L (EpcamL/L) or R80Q (EpcamQ/Q) EpCAM. Histological changes associated with a loss of HAI-2 function, including villous atrophy and tufting (A, arrowheads) in the small intestine, and increased epithelial erosion (A, open arrowheads), loss of mucin-producing goblet cells (A, arrows) and a loss of normal crypt structure in large intestines, were unaffected in mice expressing cleavage-resistant EpCAM. Similarly, loss of membrane-associated EpCAM expression (B, arrowheads) persisted in both the Vil-Cre+/0;Spint2fl/−;EpcamL/L and the Vil-Cre+/0;Spint2fl/−;EpcamQ/Q mice. In contrast, overall expression and membrane localization of claudin-7 (C, arrowheads) were largely restored in HAI-2-deficient mice expressing R80L or R80Q EpCAM. Scale bars: 50 µm (A); 25 µm (B,C). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37539662), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human EpCAM/TROP1 Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

0.25 µg/106 cells
Sample: HT‑29 human colon adenocarcinoma cell line

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed BG01V human embryonic stem cells, HT-29 Human Colon Adenocarcinoma Cell Line and Jurkat Human Acute T Cell Leukemia Cell Line cells.

Immunohistochemistry

0.3-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human colon

Simple Western

10 µg/mL
Sample: A549 human lung carcinoma cell line exosomes, HT‑29 human colon adenocarcinoma cell line exosomes, and COLO 205 human colorectal adenocarcinoma cell line whole cell lysate (WCL)

Western Blot

0.2 µg/mL
Sample: HCT‑116 human colorectal carcinoma cell line, MCF‑7 human breast cancer cell line, and BG01V human embryonic stem cells

Reviewed Applications

Read 1 review rated 5 using AF960 in the following applications:

Flow Cytometry Panel Builder

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.<br/>*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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

Alternate Names

17-1A, CD326, GA733-2, gp40, KS1/4, M4S1, TACSTD1, TROP1

Entrez Gene IDs

4072 (Human); 17075 (Mouse); 171577 (Rat); 102123042 (Cynomolgus Monkey)

Gene Symbol

EPCAM

UniProt

CAA32870

Additional EpCAM/TROP1 Products

Product Documents for Human EpCAM/TROP1 Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human EpCAM/TROP1 Antibody

For research use only

Citations for Human EpCAM/TROP1 Antibody

Customer Reviews for Human EpCAM/TROP1 Antibody (1)

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  • Human EpCAM/TROP-1 Antibody
    Name: Pauline Jeannot
    Application: Immunocytochemistry
    Sample Tested: MDA MB 231 cells
    Species: Human
    Verified Customer | Posted 11/16/2023
    MDA-MB 231 cells stained with AF960 antibody (red)
    Human EpCAM/TROP1 Antibody AF960

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