EpCAM/TROP1 Antibody (158210) [Unconjugated]

EpCAM/TROP-1 was detected in immersion fixed paraffin-embedded sections of human colon using Mouse Anti-Human EpCAM/TROP-1 Monoclonal Antibody (MAB960) at 1µg/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). Tissue was stained using the Alexa Fluor™ 647 Goat anti-Mouse IgG Secondary Antibody at 1:200 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR647MS) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the membrane. Protocol available in COMET™ Panel Builder.

EpCAM/TROP-1 was detected in immersion fixed HT-29 human colon adenocarcinoma cell line using Mouse Anti-Human EpCAM/TROP-1 Monoclonal Antibody (Catalog # MAB960) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 493-conjugated Anti-Mouse IgG Secondary Antibody (green; NL009) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

EpCAM/TROP-1 was detected in immersion fixed HT-29 human colon adenocarcinoma cell line using Mouse Anti-Human EpCAM/TROP-1 Monoclonal Antibody (Catalog # MAB960) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (yellow; NL007) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

EpCAM/TROP-1 was detected in immersion fixed paraffin-embedded sections of human adenocarcinoma using Mouse Anti-Human EpCAM/TROP-1 Monoclonal Antibody (Catalog # MAB960) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to cancer cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

EpCAM/TROP‑1 was detected in immersion fixed HT‑29 human colon adenocarcinoma cell line (positive) and Daudi human Burkitt's lymphoma cell line (negative) cells using Mouse Anti-Human EpCAM/TROP‑1 Monoclonal Antibody (Catalog # MAB960) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cell surface and cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

• Reactivity: Hu
• Applications: WB, IHC, ICC/IF, mIF
• Clone: 158210
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated

EpCAM/TROP1 Antibody (158210) [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human EpCAM/TROP-1
  Gln24-Lys265
  Accession # P16422
• Specificity: Detects human EpCAM/TROP-1 in direct ELISAs and Western blots. This antibody detects an epitope found in the extracellular domain between amino acids 136 and 265. In direct ELISAs and Western blots, no cross-reactivity with recombinant human (rh) ALCAM, rhBCAM, rhMCAM, rhNCAM-L1, or recombinant mouse (rm) OCAM is observed. In direct ELISAs, less than 5% cross-reactivity with rmEpCAM/TROP-1 is observed.
• Source: N/A
• Isotype: IgG2a
• Clonality: Monoclonal
• Host: Mouse
• Gene: EPCAM
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 8-25 ug/mL
  • Immunohistochemistry 8-25 ug/mL
  • Multiplex Immunofluorescence 3-25 ug/mL
  • Western Blot 1 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. See Certificate of Analysis for details.
  *Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Notes

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

Alternate Names for EpCAM/TROP1 Antibody (158210) [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 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).

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 (MAB960)(14)

Publications using MAB960

• K Kim, YJ Kang, SR Park, DS Kim, SW Lee, K Ko, D Ponndorf, K Ko Effect of leaf position and days post-infiltration on transient expression of colorectal cancer vaccine candidate proteins GA733-Fc and GA733-FcK in Nicotiana benthamiana plant PeerJ, 2021-04-07;9(0):e10851. 2021-04-07 [PMID: 33868796] (ELISA Capture, Western Blot, Human, Nicotiana benthamiana)
• Ye-Rin Lee, Chae-Yeon Lim, Sohee Lim, Se Ra Park, Jong-Pil Hong, Jinhee Kim et al. Expression of Colorectal Cancer Antigenic Protein Fused to IgM Fc in Chinese Cabbage (Brassica rapa) Plants (Basel) 10/30/2020 [PMID: 33143243]
• Tharagan Kumar, Ruben R. G. Soares, Leyla Ali Dholey, Harisha Ramachandraiah, Negar Abbasi Aval, Zenib Aljadi et al. Multi-layer assembly of cellulose nanofibrils in a microfluidic device for the selective capture and release of viable tumor cells from whole blood Nanoscale 11/5/2020 [PMID: 33103175]
• Ping Wei, Fei Wu, Bin Kang, Xiaohua Sun, Fabienne Heskia, Alexandre Pachot et al. Plasma extracellular vesicles detected by Single Molecule array technology as a liquid biopsy for colorectal cancer Journal of Extracellular Vesicles 9/1/2020 [PMID: 32944195]
• MA Witek, RD Aufforth, H Wang, JW Kamande, JM Jackson, SR Pullagurla, ML Hupert, J Usary, WZ Wysham, D Hilliard, S Montgomery, V Bae-Jump, LA Carey, PA Gehrig, MI Milowsky, CM Perou, JT Soper, YE Whang, JJ Yeh, G Martin, SA Soper Discrete microfluidics for the isolation of circulating tumor cell subpopulations targeting fibroblast activation protein alpha and epithelial cell adhesion molecule NPJ Precis Oncol, 2017-07-25;1(0):. 2017-07-25 [PMID: 29657983] (Cell Capture, Flow Cytometry, ICC, IHC-P, Human, Xenograft)
• MJ Mitchell, J Webster, A Chung, PP Guimarães, OF Khan, R Langer Polymeric mechanical amplifiers of immune cytokine-mediated apoptosis Nat Commun, 2017-03-20;8(0):14179. 2017-03-20 [PMID: 28317839] (Neutralization, Mouse)
• 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)
• Zhenyu Cai, Anling Zhang, Swati Choksi, Weihua Li, Tao Li, Xue-Min Zhang et al. Activation of cell-surface proteases promotes necroptosis, inflammation and cell migration Cell Research 8/1/2016 [PMID: 27444869]
• GW Krampitz, BM George, SB Willingham, JP Volkmer, K Weiskopf, N Jahchan, AM Newman, D Sahoo, AJ Zemek, RL Yanovsky, JK Nguyen, PJ Schnorr, PK Mazur, J Sage, TA Longacre, BC Visser, GA Poultsides, JA Norton, IL Weissman Identification of tumorigenic cells and therapeutic targets in pancreatic neuroendocrine tumors Proc Natl Acad Sci USA, 2016-03-31;113(16):4464-9. 2016-03-31 [PMID: 27035983] (Phagocytosis Assay, Human)
• Vin Yee Chung, Tuan Zea Tan, Ming Tan, Meng Kang Wong, Kuee Theng Kuay, Zhe Yang et al. GRHL2-miR-200-ZEB1 maintains the epithelial status of ovarian cancer through transcriptional regulation and histone modification Scientific Reports 2/18/2016 [PMID: 26887977]
• F.Q. Zhou, Y.M. Qi, H. Xu, Q.Y. Wang, X.S. Gao, H.G. Guo Expression of EpCAM and Wnt/ beta -catenin in human colon cancer Genetics and Molecular Research 5/4/2015 [PMID: 25966221]
• Zhao X, Parpart S, Takai A, Roessler S, Budhu A, Yu Z, Blank M, Zhang Y, Jia H, Ye Q, Qin L, Tang Z, Thorgeirsson S, Wang X Integrative genomics identifies YY1AP1 as an oncogenic driver in EpCAM(+) AFP(+) hepatocellular carcinoma. Oncogene, 2015-01-19;34(39):5095-104. 2015-01-19 [PMID: 25597408] (Western Blot, Human)
• Macarthur K, Kao G, Chandrasekaran S, Alonso-Basanta M, Chapman C, Lustig R, Wileyto E, Hahn S, Dorsey J Detection of brain tumor cells in the peripheral blood by a telomerase promoter-based assay. Cancer Res, 2014-02-13;74(8):2152-9. 2014-02-13 [PMID: 24525740] (IHC, Western Blot, Human, Mouse)
• Yamashita T, Budhu A, Forgues M, Wang XW Activation of hepatic stem cell marker EpCAM by Wnt-beta-catenin signaling in hepatocellular carcinoma. Cancer Res., 2007-11-15;67(22):10831-9. 2007-11-15 [PMID: 18006828] (Western Blot, Human)

FAQs for EpCAM/TROP1 Antibody (MAB960). (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:
  • P16422()