Overview

  • Product name
    Anti-EpCAM antibody [Ber-EP4]
    See all EpCAM primary antibodies
  • Description
    Mouse monoclonal [Ber-EP4] to EpCAM
  • Host species
    Mouse
  • Specificity
    Epithelial specific antigen is cell surface glycoprotein and is broadly distributed in epithelial cells and displays a highly conserved expression in carcinomas. Epithelial specific antigen has been know to play an important role as tumor-cell marker in lymph nodes from patients with esophageal carcinoma. Epithelial specific antigen can be used to distinguish between basal cell and basosquamous carcinomas and squamous cell carcinoma of the skin.
  • Tested applications
    Suitable for: IHC-FoFr, IHC-P, IHC-Fr, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    BALB/C mice were immunized with human breast carcinoma cell line, MCF-7.

  • Positive control
    • Kidney tissue sections.

Properties

Applications

Our Abpromise guarantee covers the use of ab7504 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-FoFr Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC-Fr Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.

Target

  • Function
    May act as a physical homophilic interaction molecule between intestinal epithelial cells (IECs) and intraepithelial lymphocytes (IELs) at the mucosal epithelium for providing immunological barrier as a first line of defense against mucosal infection. Plays a role in embryonic stem cells proliferation and differentiation. Up-regulates the expression of FABP5, MYC and cyclins A and E.
  • Tissue specificity
    Highly and selectively expressed by undifferentiated rather than differentiated embryonic stem cells (ESC). Levels rapidly diminish as soon as ESC's differentiate (at protein levels). Expressed in almost all epithelial cell membranes but not on mesodermal or neural cell membranes. Found on the surface of adenocarcinoma.
  • Involvement in disease
    Defects in EPCAM are the cause of diarrhea type 5 (DIAR5) [MIM:613217]. It is an intractable diarrhea of infancy characterized by villous atrophy and absence of inflammation, with intestinal epithelial cell dysplasia manifesting as focal epithelial tufts in the duodenum and jejunum.
    Defects in EPCAM are a cause of hereditary non-polyposis colorectal cancer type 8 (HNPCC8) [MIM:613244]. HNPCC is a disease associated with marked increase in cancer susceptibility. It is characterized by a familial predisposition to early-onset colorectal carcinoma (CRC) and extra-colonic tumors of the gastrointestinal, urological and female reproductive tracts. HNPCC is reported to be the most common form of inherited colorectal cancer in the Western world. Clinically, HNPCC is often divided into two subgroups. Type I is characterized by hereditary predisposition to colorectal cancer, a young age of onset, and carcinoma observed in the proximal colon. Type II is characterized by increased risk for cancers in certain tissues such as the uterus, ovary, breast, stomach, small intestine, skin, and larynx in addition to the colon. Diagnosis of classical HNPCC is based on the Amsterdam criteria: 3 or more relatives affected by colorectal cancer, one a first degree relative of the other two; 2 or more generation affected; 1 or more colorectal cancers presenting before 50 years of age; exclusion of hereditary polyposis syndromes. The term 'suspected HNPCC' or 'incomplete HNPCC' can be used to describe families who do not or only partially fulfill the Amsterdam criteria, but in whom a genetic basis for colon cancer is strongly suspected. Note=HNPCC8 results from heterozygous deletion of 3-prime exons of EPCAM and intergenic regions directly upstream of MSH2, resulting in transcriptional read-through and epigenetic silencing of MSH2 in tissues expressing EPCAM.
  • Sequence similarities
    Belongs to the EPCAM family.
    Contains 1 thyroglobulin type-1 domain.
  • Post-translational
    modifications
    Hyperglycosylated in carcinoma tissue as compared with autologous normal epithelia. Glycosylation at Asn-198 is crucial for protein stability.
  • Cellular localization
    Lateral cell membrane. Cell junction > tight junction. Co-localizes with CLDN7 at the lateral cell membrane and tight junction.
  • Information by UniProt
  • Database links
  • Alternative names
    • 17 1A antibody
    • 323/A3 antibody
    • Adenocarcinoma associated antigen antibody
    • Adenocarcinoma-associated antigen antibody
    • Antigen identified by monoclonal antibody AUA1 antibody
    • AUA1 antibody
    • CD326 antibody
    • CD326 antigen antibody
    • Cell surface glycoprotein Trop 1 antibody
    • Cell surface glycoprotein Trop 2 antibody
    • Cell surface glycoprotein Trop-1 antibody
    • CO 17A antibody
    • CO17 1A antibody
    • CO17A antibody
    • DIAR5 antibody
    • EGP 2 antibody
    • EGP antibody
    • EGP2 antibody
    • EGP314 antibody
    • EGP40 antibody
    • Ep CAM antibody
    • Ep-CAM antibody
    • EPCAM antibody
    • EPCAM_HUMAN antibody
    • EpCAM1 antibody
    • Epithelial cell adhesion molecule antibody
    • Epithelial Cell Adhesion Molecule Intracellular Domain (EpCAM-ICD) antibody
    • Epithelial cell surface antigen antibody
    • Epithelial cellular adhesion molecule antibody
    • Epithelial glycoprotein 1 antibody
    • Epithelial glycoprotein 314 antibody
    • Epithelial glycoprotein antibody
    • ESA antibody
    • GA733 1 antibody
    • GA733 2 antibody
    • GA733-2 antibody
    • gastrointestinal tumor-associated antigen 2, 35-KD glycoprotein antibody
    • gp4 antibody
    • hEGP 2 antibody
    • hEGP314 antibody
    • HNPCC8 antibody
    • Human epithelial glycoprotein 2 antibody
    • KS 1/4 antigen antibody
    • KS1/4 antibody
    • KSA antibody
    • Ly74 antibody
    • Lymphocyte antigen 74 antibody
    • M1S 1 antibody
    • M1S2 antibody
    • M4S1 antibody
    • Major gastrointestinal tumor associated protein GA733 2 antibody
    • Major gastrointestinal tumor-associated protein GA733-2 antibody
    • mEGP314 antibody
    • Membrane component chromosome 4 surface marker (35kD glycoprotein) antibody
    • Membrane component, chromosome 4, surface marker 1 antibody
    • Membrane component, chromosome 4, surface marker antibody
    • MIC18 antibody
    • MK 1 antibody
    • Protein 289A antibody
    • TACD1 antibody
    • TACSTD1 antibody
    • TROP1 antibody
    • Tumor associated calcium signal transducer 1 antibody
    • Tumor associated calcium signal transducer 2 precursor antibody
    • Tumor-associated calcium signal transducer 1 antibody
    see all

Images

  • ab7504 staining EpCAM in formalin-fixed, paraffin-embedded Human breast carcinoma tissue by Immunohistochemistry. Staining was detected using DAB.
  • Immunocytochemistry/ Immunofluorescence analysis of HepG2 cells labeling EpCAM with ab7504 at 1/200 dilution. Cells were fixed in methanol. Staining with ab7504 at 1/200 was carried out for 16 hours at 22°C in PBS buffer. ab150117, a Goat Anti-mouse IgG H&L (Alexa Fluor® 488) preadsorbed secondary antibody was used at 1/200 dilution. DAPI was used to counterstain.

    See Abreview

  • ab7504 staining EpCAM in Human liver tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% serum for 1 hour at 20°C; antigen retrieval was by heat mediation (microwave) in a commercial retrieval buffer. Samples were incubated with primary antibody (1/250 in 1% serum) for 12 hours at 4°C. A Biotin-conjugated Horse anti-mouse IgG polyclonal (1/200) was used as the secondary antibody.

    See Abreview

References

This product has been referenced in:
  • van Lith SAM  et al. The effect of subcellular localization on the efficiency of EGFR-targeted VHH photosensitizer conjugates. Eur J Pharm Biopharm 124:63-72 (2018). Human . Read more (PubMed: 29274374) »
  • Watanabe J  et al. Clinical features of squamous cell lung cancer with anaplastic lymphoma kinase (ALK)-rearrangement: a retrospective analysis and review. Oncotarget 9:24000-24013 (2018). Read more (PubMed: 29844868) »
See all 7 Publications for this product

Customer reviews and Q&As

1-10 of 17 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HepG2)
Permeabilization
No
Specification
HepG2
Fixative
Methanol

Dr. Kirk Mcmanus

Verified customer

Submitted Jan 16 2017

Application
Flow Cytometry
Sample
Human Cell (Breast cancer cell (MCF7))
Permeabilization
No
Gating Strategy
Standard
Specification
Breast cancer cell (MCF7)
Preparation
Cell harvesting/tissue preparation method: Adherent. MCF7 cell line was cultured in DMEM medium, 10% FBS. Volume used was 15 mL for 75 cm2 flask.
Sample buffer: DMEM + 10% FBS

Miss. Soledad Carinelli

Verified customer

Submitted May 20 2015

Answer


TAG72 antibody, ab16838, Lot GR99749-1: 66.6 ug/ml


Epcam antibody (Ber-EP4), ab7504, Lot GR103717-1: 47.3 ug/ml

Read More

Answer

Thank you for contacting us.

Unfortunately, this antibody has not been purified and therefore, its concentration has not been determined. However, we estimate that the antibody concentration in this tissue culture supernatant is 1-3 mg/ml.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

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Answer

Thank you for your inquiry.

For ab758, the immunogen that was used was cytoskeletal extract of human hepatocellular carcinoma cell line, Hep3B. We have not validated this antibody in pig. But the human sequence is 85% homologous with the pig cytokeratin 8 sequence. We generally say anything with >85% homology would be expected to cross-react, so this is right on the edge. However, we do have two anti-cytokeratin 8 antibodies that we know do work in pig as ab2531 and ab2530.

https://www.abcam.com/Cytokeratin-8-antibody-C-51-ab2531.html

https://www.abcam.com/Cytokeratin-8-antibody-C-43-ab2530.html

For ab27988, this antibody has been validated in pig and we therefore guarantee it will work for pig in IHC-P, IHC-Fr in accordance with our Abpromise.

For ab7504, the immunogen was human breast carcinoma cell line, MCF-7. Pig and human EpCAM is 83% homologous, so this is a little low. We have an anti-EpCAM that is predicted (>90% homology) to cross-react with pig, and that is ab71916. However, this hasn't been validated and therefore not guaranteed in pig.

https://www.abcam.com/EpCAM-antibody-ab71916.html

I have to check with the lab on ab32362 and ab124687 since I don't have the immunogens readily available to BLAST.

For ab28028, full length native protein (purified): Vimentin isolated from bovine eye lens was used as an immunogen. Pig vimentin and bovine vimentin are 97% homologous, so there is a good chance this will cross-react with pig. However this hasn't been tested and therefore isn't validated in pig. We do have a few vimentin antibodies that work in pig though,like ab20346 and ab8069.

https://www.abcam.com/Vimentin-antibody-VI-10-ab20346.html

https://www.abcam.com/Vimentin-antibody-V9-ab8069.html

For ab11333, cultured human thymic fibroblasts was used as an immunogen. We don't have a particular protein sequence to look up for this one, so unfortunately we don't know how/if it will work with pig.

I hope this information helps. I'll be in touch again once I hear about ab32362 and ab124687.

Read More

Question
Answer

Thank you for contacting us.

I was not able to find a purified, concentrated formulation of the Ber-EP4 clone in our catalogue, and we have not had it in the past.

Please do not hesitate to contact us if you need any more advice or information.
.

Read More

Question
Answer

Thank you for contacting us. As shown on the datasheet of the anti-EpCAM ab71916, www.abcam.com/ab71916, SW480 whole cell lysate is a recommended positive control. As explained on the phone, to our knowledge ab7504 has been tested in Western Blot. This monoclonal may work in this application and some optimisation may be needed. Since this antibody works in IHC, it may recognise an epitope made up of non-contiguous amino acids in their native conformation. Although the amino acids of the epitope are separated from one another in the primary sequence, they can be closer to each other in the three dimensional structure of the protein. If this hypothesis is correct, the antibody will only recognize the epitope as it exists on the surface of the folded structure. It is imperative in these circumstances to run a western blot in non-denaturing conditions, and this will be noted on the datasheet in the applications section. In general, a non-denaturing condition simply means leaving SDS out of the sample and migration buffers and not heating the samples. Certain antibodies only recognize protein in its non-reduced form i.e. in an oxidized form (particularly on cysteine residues) and the reducing agents β-mercaptoethanol and DTT must be left out of the loading buffer and migration buffer (non reducing conditions). We recommend to perform a western blot using a buffer without ionic detergents (e.g. SDS) and ideally without non-ionic detergents (e.g. Triton X-100). Cell lysis with detergent-free buffer can be achieved by mechanical shearing, often with a Dounce homogenizer or by passing cells through a syringe tip. In these cases a simple Tris buffer will suffice. Do not heat the samples. For more details I would suggest to look at the protocols book : www.abcam.com/protocolsbook or the Western Blot guide https://www.abcam.com/ps/pdf/protocols/WB-beginner.pdf . I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Question
Answer

Thank you for contacting us in regards to Anti-EpCAM antibody [Ber-EP4] (ab7504) and Anti-EpCAM antibody [HEA125] (ab46714). As discussed over the phone yesterday, we do not currently have any information regarding these antibodies having been used in Western blotting but I have seen information relating to antibodies produced from these clones having been used for this application. It therefore seems likely that they should be able to be used for this application. If you would like we could take a look at the protocol you have been using to perform Western blotting in order to see if there are any suggestions we can make to improve the likelihood of seeing the desired band. I have attached a questionnaire to this email for this purpose. However, the Invitrogen Dynabeads you have purchased work like immunoprecipitation and it may be that the antibodies from these clones are more appropriate to bind to the native epitope of the protein, not the denatured form detected in the Western blot. I hope this information has been of help.

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Question
Answer

Thank you for contacting us. THE A431, HCT116 and MCF etc express this target so lysates of these can be used as positive control. For more information about the tissue that expresses this target please check Human Protein Atlas. As discussed; SW480 cell line can also be used as positive control. However we do not have tested results of this cell line which we can forward you for comparison. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Answer

Thank you for contacting us. As discussed on phone the antibody in question can be used with ab7504. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.  

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1-10 of 17 Abreviews or Q&A

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