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    epcam-antibody-e144-ab32392.pdf

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Tags & Cell Markers Cell Type Markers Tumor Associated
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-EpCAM antibody [E144] (ab32392)

  • Datasheet
  • SDS
Reviews (19)Q&A (23)References (55)

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Western blot - Anti-EpCAM antibody [E144] (ab32392)
  • Western blot - Anti-EpCAM antibody [E144] (ab32392)
  • Western blot - Anti-EpCAM antibody [E144] (ab32392)
  • Western blot - Anti-EpCAM antibody [E144] (ab32392)
  • Anti-EpCAM antibody [E144] (ab32392)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [E144] to EpCAM
  • Suitable for: WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Anti-Integrin alpha 2 antibody [EPR17338] - Low endotoxin, Azide free (ab222377)
Knockout
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Human EPCAM knockout A-431 cell lysate (ab261708)

View more associated products

Overview

  • Product name

    Anti-EpCAM antibody [E144]
    See all EpCAM primary antibodies
  • Description

    Rabbit monoclonal [E144] to EpCAM
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WBmore details
    Unsuitable for: ICC/IF,IHC-Fr or IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HCT 116, A431, MCF7, Mouse colon tissue, and Rat colon tissue.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    E144
  • Isotype

    IgG
  • Research areas

    • Tags & Cell Markers
    • Cell Type Markers
    • Tumor Associated
    • Tags & Cell Markers
    • Cell Type Markers
    • Epi / Endo-thelial

Associated products

  • Alternative Versions

    • Anti-EpCAM antibody [E144] - BSA and Azide free (ab183178)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human EPCAM knockout A-431 cell line (ab261902)
  • KO cell lysates

    • Human EPCAM knockout A-431 cell lysate (ab261708)
  • Positive Controls

    • A-431 whole cell lysate (ab7909)
  • Related Products

    • Anti-EpCAM antibody [E144] - BSA and Azide free (ab183178)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab32392 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
WB (3)
1/1000 - 1/10000. Predicted molecular weight: 39 kDa.
Notes
WB
1/1000 - 1/10000. Predicted molecular weight: 39 kDa.
Application notes
Is unsuitable for ICC/IF,IHC-Fr or IP.

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.
  • Target information above from: UniProt accession P16422 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 4072 Human
    • Entrez Gene: 17075 Mouse
    • Entrez Gene: 171577 Rat
    • Omim: 185535 Human
    • SwissProt: P16422 Human
    • SwissProt: Q99JW5 Mouse
    • SwissProt: O55159 Rat
    • Unigene: 542050 Human
    • Unigene: 4259 Mouse
    • Unigene: 106481 Rat
    see all
  • 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

  • Western blot - Anti-EpCAM antibody [E144] (ab32392)
    Western blot - Anti-EpCAM antibody [E144] (ab32392)
    All lanes : Anti-EpCAM antibody [E144] (ab32392) at 1/1000 dilution

    Lane 1 : Wild-type HCT 116 cell lysate
    Lane 2 : EPCAM knockout HCT 116 cell lysate
    Lane 3 : A431 cell lysate
    Lane 4 : MCF7 cell lysate
    Lane 5 : HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 39 kDa
    Observed band size: 37-45 kDa why is the actual band size different from the predicted?



    False colour image of Western blot: Anti-EpCAM antibody [E144] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab32392 was shown to bind specifically to EpCAM. A band was observed at 37/45 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in EPCAM knockout cell line ab281596 (knockout cell lysate ab282948). To generate this image, wild-type and EPCAM knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

  • Western blot - Anti-EpCAM antibody [E144] (ab32392)
    Western blot - Anti-EpCAM antibody [E144] (ab32392)
    All lanes : Anti-EpCAM antibody [E144] (ab32392) at 1/1000 dilution

    Lane 1 : Mouse colon tissue lysate
    Lane 2 : Rat small intestine tissue lysate
    Lane 3 : Rat colon tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size: 39 kDa

  • Western blot - Anti-EpCAM antibody [E144] (ab32392)
    Western blot - Anti-EpCAM antibody [E144] (ab32392)
    All lanes : Anti-EpCAM antibody [E144] (ab32392) at 1/2000 dilution

    Lane 1 : Wild-type A431 cell lysate
    Lane 2 : EPCAM knockout A431 cell lysate
    Lane 3 : MCF7 cell lysate
    Lane 4 : HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 39 kDa
    Observed band size: 40 kDa why is the actual band size different from the predicted?



    Lanes 1 - 4: Merged signal (red and green). Green - ab32392 observed at 40 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.

    ab32392 was shown to react with EpCAM in wild-type A431 cells in western blot with loss of signal observed in EpCAM knockout sample. Wild-type and EpCAM knockout A431 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab32392 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at a 1 in 2000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-EpCAM antibody [E144] (ab32392)
    Western blot - Anti-EpCAM antibody [E144] (ab32392)
    Anti-EpCAM antibody [E144] (ab32392) at 1/2500 dilution + A431 cell lysate

    Predicted band size: 39 kDa

  • Anti-EpCAM antibody [E144] (ab32392)
    Anti-EpCAM antibody [E144] (ab32392)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (55)

Publishing research using ab32392? Please let us know so that we can cite the reference in this datasheet.

ab32392 has been referenced in 55 publications.

  • Zhang PD  et al. Feasibility of primary human cell cultures as a model for adamantinomatous craniopharyngioma research: Evidence from RNA-Seq analysis. Oncol Lett 19:2346-2354 (2020). PubMed: 32194734
  • Jia N  et al. The Human Lung Glycome Reveals Novel Glycan Ligands for Influenza A Virus. Sci Rep 10:5320 (2020). PubMed: 32210305
  • Das B  et al. Congenital Tufting Enteropathy-Associated Mutant of Epithelial Cell Adhesion Molecule Activates the Unfolded Protein Response in a Murine Model of the Disease. Cells 9:N/A (2020). PubMed: 32290509
  • Karagonlar ZF  et al. A Novel Function for KLF4 in Modulating the De-differentiation of EpCAM-/CD133- nonStem Cells into EpCAM+/CD133+ Liver Cancer Stem Cells in HCC Cell Line HuH7. Cells 9:N/A (2020). PubMed: 32408542
  • Steins A  et al. High-grade mesenchymal pancreatic ductal adenocarcinoma drives stromal deactivation through CSF-1. EMBO Rep 21:e48780 (2020). PubMed: 32173982
View all Publications for this product

Customer reviews and Q&As

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1-10 of 19 Abreviews

IHC - Wholemount abreview for Anti-EpCAM antibody [E144]

Excellent
Abreviews
Abreviews
abreview image
Application
IHC - Wholemount
Sample
Mouse Tissue (small intestine organoids)
Specification
small intestine organoids
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Mar 24 2017

Immunocytochemistry/ Immunofluorescence abreview for Anti-EpCAM antibody [E144]

Excellent
Abreviews
Abreviews
This product is known to not work in this application or species.
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Adenocarcinoma cells)
Permeabilization
Yes - 0.1% TX-100
Specification
Adenocarcinoma cells
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jan 09 2017

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-EpCAM antibody [E144]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Pig Tissue sections (Lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Sodium Citrate pH6
Permeabilization
No
Specification
Lung
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% normal buffered formalin
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Aug 25 2016

Immunohistochemistry (Frozen sections) abreview for Anti-EpCAM antibody [E144]

Good
Abreviews
Abreviews
This product is known to not work in this application or species.
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (stomach)
Permeabilization
No
Specification
stomach
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
Acetone
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Aug 22 2016

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-EpCAM antibody [E144]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (skin tumor)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Sodium Citrate pH6
Permeabilization
No
Specification
skin tumor
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% normal buffered formalin
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Aug 15 2016

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-EpCAM antibody [E144]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (tumor)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Sodium Citrate buffer pH6
Permeabilization
No
Specification
tumor
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jul 11 2016

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-EpCAM antibody [E144]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Ovarian Tumor)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH 6
Permeabilization
No
Specification
Ovarian Tumor
Blocking step
Peroxidase Blocking Solution + UV block as blocking agent for 10 minute(s) · Concentration: 100% · Temperature: 25°C
Fixative
58°C - 1h
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jun 02 2016

ELISA abreview for Anti-EpCAM antibody [E144]

Excellent
Abreviews
Abreviews
abreview image
Application
ELISA
Sample
Human Cell (Cancer cell line)
Specification
Cancer cell line
Blocking step
PBS as blocking agent for 30 minute(s) · Concentration: 15µg/mL · Temperature: 37°C
Type
Direct
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Sep 18 2015

Western blot abreview for Anti-EpCAM antibody [E144]

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Colon cancer cell lines)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
20 µg
Specification
Colon cancer cell lines
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 27°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jun 30 2015

Immunocytochemistry/ Immunofluorescence abreview for Anti-EpCAM antibody [E144]

Poor
Abreviews
Abreviews
This product is known to not work in this application or species.
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Permeabilization
Yes - 0.5% Triton X100 in PBS
Specification
HeLa
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Kirk Mcmanus

Verified customer

Submitted Apr 04 2014

1-10 of 19 Abreviews

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