EpCAM/TROP1 Antibody (AUA1) - BSA Free

Immunohistochemistry-Paraffin: EpCAM/TROP1 Antibody (AUA1) [NB600-1182] - Analysis of a FFPE human breast carcinoma tissue section using 1:100 dilution of EpCAM/TROP1 antibody (clone AUA1) on a Bond Rx autostainer (Leica Biosystems). The assay involved 20 minutes of heat induced antigen retrieval (HIER) with 10mM sodium citrate buffer (pH 6.0) and endogenous peroxidase quenching using peroxide block. The sections were incubated with primary antibody for 30 minutes. Bond Polymer Refine Detection (Leica Biosystems) and DAB were used for signal detection which followed counterstaining with hematoxylin. Whole slide scanning and capturing of representative images (20X) were performed using Aperio AT2 (Leica Biosystems). This EpCAM antibody generated an expected immunostaining of EpCAM (CD326) protein in the membranes of the cancer cells and the inter-cellular spaces. The tumor stroma and the stromal cells did not show EpCAM immunopositivity.

Western Blot: EpCAM/TROP1 Antibody (AUA1) [NB600-1182] - Detection of EpCAM/CD326 in two Human mammary tumor cell lines, MDA-MB-231 (mesenchyme-like) and MCF-7 (epithelium-like). Dilution: 1:500 in PBS with 5% BSA. Secondary Ab: anti-Mouse IgG 1:5,000. This image was submitted via customer Review.

Immunocytochemistry/Immunofluorescence: EpCAM/TROP1 Antibody (AUA1) [NB600-1182] - A431 cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.05% Triton X-100. The cells were incubated with anti-EpCAM/TROP1 (AUA1) conjugated to Alexa Fluor 488 [NB600-1182AF488] at 10ug/ml for 1 hour at room temperature. Nuclei were counterstained with DAPI (blue). Cells were imaged using a 40X objective.

Immunohistochemistry-Paraffin: EpCAM/TROP1 Antibody (AUA1) [NB600-1182] - Analysis of a FFPE human breast carcinoma tissue with 1:500 dilution of EpCAM/TROP1 antibody (clone AUA1) on a Bond Rx autostainer (Leica Biosystems). The assay involved 20 minutes of heat induced antigen retrieval (HIER) with 10mM sodium citrate buffer (pH 6.0) and endogenous peroxidase quenching using peroxide block. The sections were incubated with primary antibody for 30 minutes. Bond Polymer Refine Detection (Leica Biosystems) and DAB were used for signal detection which followed counterstaining with hematoxylin. Whole slide scanning and capturing of representative images (20X) were performed using Aperio AT2 (Leica Biosystems). The staining was primarily localized to the membranes and inter-cellular spaces of the cancer cells. Staining intensity for 1:500 dilution was lower than what was seen in sections tested with 1:100 dilution. Staining was performed by Histowiz.

Flow Cytometry: EpCAM/TROP1 Antibody (AUA1) [NB600-1182] - Using the PE direct conjugate A cell surface stain was performed on HT-29 cells with EpCAM (AUA1) antibody NB600-1182PE (blue) and a matched isotype control NBP2-27287PE (orange). Cells were incubated in an antibody dilution of 5 ug/mL for 20 minutes at room temperature. Both antibodies were conjugated to phycoerythrin.

Western Blot: EpCAM/TROP1 Antibody (AUA1) [NB600-1182] - Analysis of A431 lysate (A) and Hek293 lysate (B) using EpCAM/CD326 antibody (AUA1) at 2 ug/ml. Non-reduced lysate was used in each lane.

Flow Cytometry: EpCAM/TROP1 Antibody (AUA1) [NB600-1182] - Intracellular flow cytometric staining of 1 x 10^6 CHO (A) and HeLa (B) cells using EpCAM/CD326 antibody (blue) and matched Isotype control (orange). An antibody concentration of 1 ug/1x10^6 cells was used.

• Reactivity: Hu
• Applications: WB, ELISA, Flow, ICC/IF, IHC, CyTOF-ready
• Clone: AUA1
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1.0 mg/ml

EpCAM/TROP1 Antibody (AUA1) - BSA Free Summary

• Description: Novus Biologicals Mouse EpCAM/TROP1 Antibody (AUA1) - BSA Free (NB600-1182) is a monoclonal antibody validated for use in IHC, WB, ELISA, Flow and ICC/IF. Anti-EpCAM/TROP1 Antibody: Cited in 12 publications. All Novus Biologicals antibodies are covered by our 100% guarantee.
• Immunogen: LoVo cell line preparation (Human).
• Localization: Cell Membrane
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: EPCAM
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • ELISA 1:100-1:2000
  • Flow Cytometry 1 ug per million cells
  • Immunocytochemistry/ Immunofluorescence 1:10-1:500. Use reported in scientific literature (PMID 21763624)
  • Immunohistochemistry 1:10-1:500
  • Immunohistochemistry-Paraffin 1:10-1:500
  • Western Blot 1:500
• Application Notes: This antibody is CyTOF ready. Flow Cytometry reported by Alvarez et al - Conference on Retroviruses and Opportunistic Infections - CROI 2014 (Abstract: The NCOR2-Nurr1-CoREST Transrepression Axis Impairs HIV Reactivation in Latently Infected Microglial Cells)
• Theoretical MW: 35 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.05% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Protein G purified

Alternate Names for EpCAM/TROP1 Antibody (AUA1) - BSA Free

• 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 cell adhesion molecule (EpCAM), also known as TROP1 and CD326, is a type I transmembrane glycoprotein that plays a role in a number of cellular processes including adhesion, signaling, maintaining stemness, proliferation, migration, and invasion (1,2). The human EpCAM protein is 314 amino acids (aa) in length with a theoretical molecular weight (MW) of ~35 kDa (1,3,4). The EpCAM protein includes a signal peptide, an extracellular N-terminal domain, a thyroglobulin type-1 domain, a carboxyl-terminal domain, a single-pass transmembrane domain, and an intracellular domain (1,3). The protein is highly conserved and has approximately 81% aa sequence identity between human and mouse (5). Trophoblast cell-surface antigen 2 (TROP2) also shares ~67% aa sequence similarity with EpCAM (1,5). While both EpCAM and TROP2 are cell surface markers expressed in the epithelium, their expression levels typically do not correlate (5). EpCAM is expressed in normal epithelial tissue and elevated expression is observed in many epithelial carcinomas (1-3) Patient tumor samples with increased EpCAM expression have been associated with poor prognosis (1). Given EpCAM's elevated expression in tumors, it has become a potential target for cancer therapy approaches, including monoclonal antibody treatment and anti-EpCAM trispecific antibodies (1,5).

EpCAM functions as an intracellular signaling molecule and contributes to regulation of epithelial-to-mesenchymal (EMT) transition (4,5). EpCAM is cleaved during the process of regulated intramembrane proteolysis (RIP) (4,5). Initial cleavage occurs at the membrane via a disintegrin and metalloprotease 17 (ADAM17) which releases EpCAM's extracellular domain (EpEx) (4,5). A secondary cleavage is mediated by presenilin 2 (PSEN2) which releases EpCAM's cytoplasmic trail (EpICD) (4,5). EpICD translocates to the nucleus where it associates with beta-catenin, FHL-2, and LEF-1 and induces transcription of genes related to EMT and tumor growth (4,5). EpCAM has also been shown to regulate structure and functionality of the apical junction complex of cells through direct interaction with claudin-7 and association with E-cadherin (2,5). Loss of EpCAM disrupts adherins junction and tight junction structure and function (2).

References

1. Mohtar MA, Syafruddin SE, Nasir SN, Low TY. Revisiting the Roles of Pro-Metastatic EpCAM in Cancer. Biomolecules. 2020; 10(2):255. https://doi.org/10.3390/biom10020255

2. Huang L, Yang Y, Yang F, et al. Functions of EpCAM in physiological processes and diseases (Review). Int J Mol Med. 2018; 42(4):1771-1785. https://doi.org/10.3892/ijmm.2018.3764

3. Brown TC, Sankpal NV, Gillanders WE. Functional Implications of the Dynamic Regulation of EpCAM during Epithelial-to-Mesenchymal Transition. Biomolecules. 2021; 11(7):956. https://doi.org/10.3390/biom11070956

4. Uniprot (P16422)

5. Schnell U, Cirulli V, Giepmans BN. EpCAM: structure and function in health and disease. Biochim Biophys Acta. 2013; 1828(8):1989-2001. https://doi.org/10.1016/j.bbamem.2013.04.018

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 (NB600-1182)(12)

Publications using NB600-1182

• Sherif, O;Khelwatty, SA;Bagwan, I;Seddon, AM;Dalgleish, A;Mudan, S;Modjtahedi, H; Expression of EGFRvIII and its co?expression with wild?type EGFR, or putative cancer stem cell biomarkers CD44 or EpCAM are associated with poorer prognosis in patients with hepatocellular carcinoma Oncology reports 2024-12-01 [PMID: 39450530]
• Titto Augustine, Peter John, Tyler Friedman, Jeeshan Jiffry, Hillary Guzik, Rifat Mannan, Riya Gupta, Catherine Delano, John M. Mariadason, Xingxing Zang, Radhashree Maitra, Sanjay Goel Potentiating effect of reovirus on immune checkpoint inhibition in microsatellite stable colorectal cancer Frontiers in Oncology 2022-10-25 [PMID: 36387154]
• Fan H, Atiya HI, Wang Y Et al. Epigenomic Reprogramming toward Mesenchymal-Epithelial Transition in Ovarian-Cancer-Associated Mesenchymal Stem Cells Drives Metastasis Cell Rep 2020-12-09 [PMID: 33296650]
  
  Details:
  Citation using the Alexa Fluor 405 version of this antibody.
• Chang ZM, Wang Z, Shao D et al. Shape Engineering Boosts Magnetic Mesoporous Silica Nanoparticle-Based Isolation and Detection of Circulating Tumor Cells. ACS Appl Mater Interfaces 2018-03-20 [PMID: 29468874] (FLOW)
  
  Details:
  This citation used the PE version of this antibody.
• Albarenque SM, Zwacka RM, Mohr A. Both human and mouse mesenchymal stem cells promote breast cancer metastasis Stem Cell Res 2011-09-01 [PMID: 21763624]
• Aleksic T, Chitnis MM, Perestenko OV et al. Type 1 insulin-like growth factor receptor translocates to the nucleus of human tumor cells. Cancer Res. 2010-08-15 [PMID: 20710042] (WB, Human)
• Ntouroupi TG, Ashraf SQ, Mcgregor SB et al. Detection of circulating tumour cells in peripheral blood with an automated scanning fluorescence microscope. Br J Cancer. 2008-09-02 [PMID: 18682708] (ICC/IF, Human)
• Conaghan P, Ashraf S, Tytherleigh M et al. Targeted killing of colorectal cancer cell lines by a humanised IgG1 monoclonal antibody that binds to membrane-bound carcinoembryonic antigen. Br J Cancer. 2008-04-08 [PMID: 18349843] (FLOW, Human)
• El-Hariry I, Pignatelli M, Lemoine NR et al. FGF-1 and FGF-2 modulate the E-cadherin/catenin system in pancreatic adenocarcinoma cell lines. Br J Cancer. 2001-06-15 [PMID: 11401320]
• Jordinson M, El-Hariry I, Calnan D et al. Vicia faba agglutinin, the lectin present in broad beans, stimulates differentiation of undifferentiated colon cancer cells. Gut. 1999-05-01 [PMID: 10205210] (WB, Human)
• Anagnostaki E, Skaarlos D, Tamvakis N et al. Immunohistochemical and immunocytochemical study of bladder carcinomas using the epithelium-specific, tumour-associated monoclonal antibodies HMFG1 and AUA1. Br J Cancer Suppl. 1990-07-01 [PMID: 2200496] (ICC/IF, IF/IHC, Human)
• Kocjan G, Sweeney E, Miller KD, Bobrow L et al. AUA1: new immunocytochemical marker for detecting epithelial cells in body cavity fluids. J Clin Pathol. 1992-04-01 [PMID: 1577978] (ICC/IF, Human)

Review for EpCAM/TROP1 Antibody (NB600-1182) (1)

Average Rating: 5

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Verified Customer WB Human 03/06/2017

Summary

• Application: Western Blot
• Sample Tested: Human breast cancer cell lines
• Species: Human
• Lot: A-3

FAQs for EpCAM/TROP1 Antibody (NB600-1182). (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
• Entrez:
  • 4072(Human)