EpCAM/TROP1 Antibody [Unconjugated]

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.

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 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 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 (Catalog # 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 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 (Catalog # 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.

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.

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.

• Reactivity: Hu
• Applications: WB, Simple Western, Flow, IHC, CyTOF-ready, ICC/IF
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

EpCAM/TROP1 Antibody [Unconjugated] Summary

• 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.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: EPCAM
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Flow Cytometry 0.25 ug/10^6 cells
  • Immunocytochemistry 5-15 ug/mL
  • Immunohistochemistry 0.3-15 ug/mL
  • Simple Western 10 ug/mL
  • Western Blot 0.2 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for EpCAM/TROP1 Antibody [Unconjugated]

• 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

Background

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).

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for EpCAM/TROP1 Antibody (AF960)(15)

Publications using AF960

• Zhao, C;Wang, D;Li, Z;Zhang, Z;Xu, Y;Liu, J;Lei, Q;Han, D;Huo, Y;Liu, S;Li, L;Zhang, Y; IL8 derived from macrophages inhibits CD8+ T-cell function by downregulating TIM3 expression through IL8-CXCR2 axis in patients with advanced colorectal cancer International immunopharmacology 2023-06-16 [PMID: 37331296] (IHC, ICC, Human)
• Matkovic Leko, I;Schneider, RT;Thimraj, TA;Schrode, N;Beitler, D;Liu, HY;Beaumont, K;Chen, YW;Snoeck, HW; A distal lung organoid model to study interstitial lung disease, viral infection and human lung development Nature protocols 2023-05-10 [PMID: 37165073] (IHC, Human)
• IM Leko, N Schrode, J Torres, M Pezet, TA Thimraj, KG Beaumont, HW Snoeck Generation and expansion of transitional lung organoids from human pluripotent stem cells bioRxiv : the preprint server for biology, 2023-02-01;0(0):. 2023-02-01 [PMID: 36778291] (ICC, Human)
• JJ Wang, N Sun, YT Lee, M Kim, T Vagner, K Rohena-Riv, Z Wang, Z Chen, RY Zhang, J Lee, C Zhang, H Tang, J Widjaja, TX Zhang, D Qi, PC Teng, YJ Jan, KC Hou, C Hamann, HM Sandler, TJ Daskivich, DJ Luthringer, NA Bhowmick, R Pei, S You, DD Vizio, HR Tseng, JF Chen, Y Zhu, EM Posadas Prostate cancer extracellular vesicle digital scoring assay - a rapid noninvasive approach for quantification of disease-relevant mRNAs Nano Today, 2023-01-03;48(0):. 2023-01-03 [PMID: 36711067] (Capture Assay, Human)
• H Yang, Q Deng, T Ni, Y Liu, L Lu, H Dai, H Wang, W Yang Targeted Inhibition of LPL/FABP4/CPT1 fatty acid metabolic axis can effectively prevent the progression of nonalcoholic steatohepatitis to liver cancer International journal of biological sciences, 2021-10-11;17(15):4207-4222. 2021-10-11 [PMID: 34803493] (IHC, Mouse)
• J Zhang, LTH Nguyen, R Hickey, N Walters, X Wang, KJ Kwak, LJ Lee, AF Palmer, E Reátegui Immunomagnetic sequential ultrafiltration (iSUF) platform for enrichment and purification of extracellular vesicles from biofluids Scientific Reports, 2021-04-13;11(1):8034. 2021-04-13 [PMID: 33850163] (Flow Cytometry, ICC, Human)
• KM Chan, J Gleadle, J Li, TD Michl, K Vasilev, M MacGregor Improving hexaminolevulinate enabled cancer cell detection in liquid biopsy immunosensors Scientific Reports, 2021-03-31;11(1):7283. 2021-03-31 [PMID: 33790357] (ELISA Capture, Human)
• LL Lin, ER Kost, CL Lin, P Valente, CM Wang, MG Kolonin, AC Daquinag, X Tan, N Lucio, CN Hung, CP Wang, NB Kirma, TH Huang PAI-1-Dependent Inactivation of SMAD4-Modulated Junction and Adhesion Complex in Obese Endometrial Cancer Cell Rep, 2020-10-13;33(2):108253. 2020-10-13 [PMID: 33053339] (CyTof, Human)
• N Sun, YT Lee, RY Zhang, R Kao, PC Teng, Y Yang, P Yang, JJ Wang, M Smalley, PJ Chen, M Kim, SJ Chou, L Bao, J Wang, X Zhang, D Qi, J Palomique, N Nissen, SB Han, S Sadeghi, RS Finn, S Saab, RW Busuttil, D Markovic, D Elashoff, HH Yu, H Li, AP Heaney, E Posadas, S You, JD Yang, R Pei, VG Agopian, HR Tseng, Y Zhu Purification of HCC-specific extracellular vesicles on nanosubstrates for early HCC detection by digital scoring Nat Commun, 2020-09-07;11(1):4489. 2020-09-07 [PMID: 32895384] (ELISA Capture, Human)
• S Yoshida, H Miwa, T Kawachi, S Kume, K Takahashi Generation of intestinal organoids derived from human pluripotent stem cells for drug testing Sci Rep, 2020-04-06;10(1):5989. 2020-04-06 [PMID: 32249832] (ICC, Human)
• D Garnier, R Li, F Delbos, A Fourrier, C Collet, C Guguen-Gui, C Chesné, TH Nguyen Expansion of human primary hepatocytes in vitro through their amplification as liver progenitors in a 3D organoid system Sci Rep, 2018-05-29;8(1):8222. 2018-05-29 [PMID: 29844473] (ICC, Human)
• Fast and selective cell isolation from blood sample by microfiber fabric system with vacuum aspiration Sci Technol Adv Mater, 2016-11-25;17(1):807-815. 2016-11-25 [PMID: 27933120] (Cell Selection, Human)
• HER2 and EGFR overexpression support metastatic progression of prostate cancer to bone Cancer Res., 2016-10-28;0(0):. 2016-10-28 [PMID: 27793843] (CTC, Human)
• EpCAM-Regulated Transcription Exerts Influences on Nanomechanical Properties of Endometrial Cancer Cells that Promote Epithelial-to-Mesenchymal Transition Cancer Res, 2016-08-28;0(0):. 2016-08-28 [PMID: 27569206] (Adhesion Assay, Human)
• 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 Stromal gene expression defines poor-prognosis subtypes in colorectal cancer. Nat Genet, 2015-02-23;47(4):320-9. 2015-02-23 [PMID: 25706628] (IHC, Human)

Review for EpCAM/TROP1 Antibody (AF960) (1)

Average Rating: 5

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Pauline Jeannot ICC Human 11/16/2023

Summary

• Application: Immunocytochemistry
• Sample Tested: MDA MB 231 cells
• Species: Human
• Lot: FOX0821051

FAQs for EpCAM/TROP1 Antibody (AF960). (Showing 1 - 1 of 1 FAQs).

1. I'm looking for EpCAM antibody for rat. Do you have any antibody which can use FACS?
   • We unfortunately do not have any EpCAM antibodies validated in FACS for detection of the rat protein. I am sorry for any inconvenience. I do have three products to EpCAM that can detect the rat protein. These products have been validated to detect the native rat protein, so they may work for FACS however they have not yet been tested in that particular application. If you would like to try them in your experiment you would qualify for our Innovator's Reward Program. This program was designed to help scientists with the cost of their antibodies in return for valuable information. If you decide to test an antibody in a previously untested species or application we will issue you a credit for the purchase price of the antibody in exchange for a review of your experiment using our product.

Bioinformatics

• Gene Symbol: EPCAM
• Uniprot:
  • CAA32870()