EpCAM/TROP1 Antibody (AUA1) - BSA Free

Novus Biologicals | Catalog # NB600-1182

Novus Biologicals
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Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, ELISA, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, CyTOF-ready

Cited:

Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, IF/IHC

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # AUA1

Format

BSA Free
View all EpCAM/TROP1 Primary Antibodies »

Product Specifications

Immunogen

LoVo cell line preparation (Human).

Localization

Cell Membrane

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

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.

Scientific Data Images for EpCAM/TROP1 Antibody (AUA1) - BSA Free

Immunohistochemistry-Paraffin: EpCAM/TROP1 Antibody (AUA1) - BSA Free [NB600-1182]

Immunohistochemistry-Paraffin: EpCAM/TROP1 Antibody (AUA1) - BSA Free [NB600-1182]

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)BSA Free [NB600-1182]

Western Blot: EpCAM/TROP1 Antibody (AUA1)BSA Free [NB600-1182]

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) - BSA Free [NB600-1182]

Immunocytochemistry/ Immunofluorescence: EpCAM/TROP1 Antibody (AUA1) - BSA Free [NB600-1182]

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) - BSA Free [NB600-1182]

Immunohistochemistry-Paraffin: EpCAM/TROP1 Antibody (AUA1) - BSA Free [NB600-1182]

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) - BSA Free [NB600-1182]

Flow Cytometry: EpCAM/TROP1 Antibody (AUA1) - BSA Free [NB600-1182]

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)BSA Free [NB600-1182]

Western Blot: EpCAM/TROP1 Antibody (AUA1)BSA Free [NB600-1182]

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) - BSA Free [NB600-1182]

Flow Cytometry: EpCAM/TROP1 Antibody (AUA1) - BSA Free [NB600-1182]

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.

Applications for EpCAM/TROP1 Antibody (AUA1) - BSA Free

Application
Recommended Usage

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)

Reviewed Applications

Read 1 review rated 5 using NB600-1182 in the following applications:

Flow Cytometry Panel Builder

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS

Format

BSA Free

Preservative

0.05% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: EpCAM/TROP1

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

Long Name

Epithelial Cell Adhesion Molecule

Alternate Names

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

Entrez Gene IDs

4072 (Human)

Gene Symbol

EPCAM

Additional EpCAM/TROP1 Products

Product Documents for EpCAM/TROP1 Antibody (AUA1) - BSA Free

Certificate of Analysis

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Product Specific Notices for EpCAM/TROP1 Antibody (AUA1) - BSA Free

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.

Citations for EpCAM/TROP1 Antibody (AUA1) - BSA Free

Customer Reviews for EpCAM/TROP1 Antibody (AUA1) - BSA Free (1)

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Customer Images

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  • EpCAM/TROP1 Antibody (AUA1)
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Human breast cancer cell lines
    Species: Human
    Verified Customer | Posted 03/06/2017
    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.
    EpCAM/TROP1 Antibody (AUA1) - BSA Free NB600-1182

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FAQs for EpCAM/TROP1 Antibody (AUA1) - BSA Free

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  • Q: I'm looking for EpCAM antibody for rat. Do you have any antibody which can use FACS?

    A:

    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.

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