Overview

  • Product name
    Anti-EpCAM antibody [AUA1]
    See all EpCAM primary antibodies
  • Description
    Mouse monoclonal [AUA1] to EpCAM
  • Host species
    Mouse
  • Tested applications
    Suitable for: ICC/IF, Flow Cyt, ELISA, IHC-Pmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Tissue, cells or virus corresponding to EpCAM. LoVo cell line preparation (Human).

  • Positive control
    • Flow Cyt: DU 145 cells. ICC/IF: H358 cells. IHC-P: Human breast carcinoma tissue.
  • General notes

    This antibody can be used to detect epithelial cells.

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: PBS, 50% Glycerol
  • Concentration information loading...
  • Purity
    Protein G purified
  • Primary antibody notes
    Cross reacts with breast carcinomas, kidney tubules, tonsil crypt epithelium and cells in some serious effusions. Useful in characterisation of all epithelial cells.
  • Clonality
    Monoclonal
  • Clone number
    AUA1
  • Myeloma
    P3-x63-Ag8
  • Isotype
    IgG1
  • Light chain type
    kappa
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab20160 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
ICC/IF Use at an assay dependent concentration. Used at a dilution of 1/250 for 1 hr - see Abreview for further information.
Flow Cyt Use 1µg for 106 cells.

(methanol fixed cells)

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

ELISA Use at an assay dependent concentration.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

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

  • Validation of Integrin α6/CD49f to identify CRC cells in patient samples.

    Freshly harvested tumor or normal tissue was snap frozen and banked at −80°C. A gastrointestinal pathologist confirmed the histopathology diagnosis of each specimen independently. Normal tissue was obtained from a site distal from the primary colon tumor. Fresh frozen tissues were sectioned at a 6 µm thickness. Slides were fixed with 4% paraformaldehyde, air-dried, and stored at −20°C until use. After treatment with 10% normal goat serum and 0.1% Triton X-100 for 45 min, slides were incubated with monoclonal affinity purified mouse anti-human EpCAM (ab20160, Abcam) at a final dilution of 1∶200.

    Panels shown are EpCAM (ab20160, 1/200 dilution) and merge for normal tissue.

  • Overlay histogram showing DU 145 (Human prostate carcinoma cell line) cells stained with ab20160 (red line).

    The cells were fixed with methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab20160, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

    This antibody gave a slightly decreased signal in DU 145 cells fixed with 4% paraformaldehyde (10 min) used under the same conditions.

    Please note that Abcam does not have data for use of this antibody on non-fixed cells. We welcome any customer feedback.

  • ab20160 at a 1/250 dilution staining human non small cell lung cancer cells (H358 cells) by immunocytochemistry.

    The antibody was incubated with the cells for 1 hour and then detected using an Alexa-Fluor® 546 conjugated goat anti mouse antibody.

    This image is courtesy of an Abreview submitted by an anonymous researcher on 30 January 2006.

    See Abreview

  • IHC image of ab20160 staining in human breast carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab20160, 10µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with hematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

References

This product has been referenced in:
  • Zhang C  et al. TGFß1 Promotes Breast Cancer Local Invasion and Liver Metastasis by Increasing the CD44high/CD24- Subpopulation. Technol Cancer Res Treat 17:1533033818764497 (2018). Read more (PubMed: 29658391) »
  • Wei JC  et al. Tumor-associated Lymphatic Endothelial Cells Promote Lymphatic Metastasis By Highly Expressing and Secreting SEMA4C. Clin Cancer Res 23:214-224 (2017). Read more (PubMed: 27401250) »
See all 12 Publications for this product

Customer reviews and Q&As

1-10 of 11 Abreviews or Q&A

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Brain)
Permeabilization
Yes - Triton X-100
Specification
Brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Dec 12 2016

Application
Flow Cytometry
Sample
Human Cell (Hes3 Stem cell)
Permeabilization
No
Specification
Hes3 Stem cell

Abcam user community

Verified customer

Submitted Oct 21 2015

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cultured Cells (breast cancer cell line(MCF-7))
Permeabilization
No
Specification
breast cancer cell line(MCF-7)
Fixative
Formaldehyde

Mr. Chang Su Jeon

Verified customer

Submitted Aug 20 2015

Question
Answer


Unfortunately, we do not sell ab20160 conjugated to biotin. However, we do sell a kit: EasyLink Biotin (type A) Conjugation Kit (3 x 100µg Biotin) (ab102864) which allows you to conjugate ab20160 to biotin yourself which may be of interest to you.

Read More

Answer

Thank you for your enquiry and interest in our products.

I can confirm that this particular antibody has not yet been tested in Western blot (WB) application so we do not have do not have data to share with our customers. We have other antibodies against this target (EpCAM) which have been characterized for WB.

https://www.abcam.com/ab8666:

https://www.abcam.com/ab32392:

https://www.abcam.com/ab8601:

The expected band size of the EpCAM is around 34,932 kDa, however the mature processed protein may have higher MW due to N-glycosylation. You may wish to take a look at this Swiss-Prot site for further information about the potencial post-translational modification:

http://www.uniprot.org/uniprot/P16422

If you need any further assistance in the future, please do not hesitate to contact me.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
ELISA
Sample
Human Cell (Circullating Tumor Cells)
Specification
Circullating Tumor Cells
Blocking step
Serum as blocking agent for 25 minute(s) · Concentration: 2µg/mL · Temperature: 25°C
Type
Other

Mr. Gabriel Ortega

Verified customer

Submitted Dec 20 2012

Question
Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement of ab20160.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Read More

Answer

Thank you for contacting us.

As ab20160, ab89992, ab8666 have as yet not been tested with mouse samples, I would therefore not be able to say if they would cross react with the mouse tissue or not. I think a different antibody which may be more worthwhile trying is ab124825. This has been tested in mouse and found not to react, and as it is a monolconal, this reactivity is more likely to be maintained through different lots.

This antibody has only been tested in WB, IHC-P andICC. If your customer would be interested in participating in the testing discount scheme to use the antibody in IHC-Fr and IF then I would be able to generate a discount code for them. If they would be interested in this I would need their email address and the institutethey are from.

More information on the testing discount scheme can be found from the following link:

https://www.abcam.com/collaborationdiscount

I hope this information has been of help. If you have any further questions, please do not hesitate to ask.

Read More

Question
Answer

Thank you for your telephone call on Friday.

As discussed on the telephone, we recommend not to dilute before storing as antibodies not stable at lower concentrations. I can suggest to aliquot and freezewithout diluting in 10 ul aliquots (not less as this may evaporate in the freezer and affect the antibody).

I can confirm thatlot GR41981 of ab20160 has concentration of 1.1mg/ml.Therefore, if there is the equivalent of 100 ug provided in the vial, there should be approximately 90 ul in the vial. The vial may need to be spun in the microcentrifuge to ensure all the liquid has come down from the lid.

I hope this will be helpful. Please do not hesitate to let me know if you have less than 90 ul in the original vial that was sent.

Read More

Question
Answer

Thank you for contacting us.

I am sorry that the vial you received did not contain the full amount of the product and I apologize for the inconvenience. I have issued a free of charge replacement vial.

Please do not hesitate to contact us if you need anything further.

Read More

1-10 of 11 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Sign up