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  1. Link

    epcam-antibody-4a7-c-terminal-ab224826.pdf

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

Anti-EpCAM antibody [4A7] - C-terminal (ab224826)

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  • SDS
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EpCAM antibody [4A7] - C-terminal (ab224826)
  • Western blot - Anti-EpCAM antibody [4A7] - C-terminal (ab224826)
  • Western blot - Anti-EpCAM antibody [4A7] - C-terminal (ab224826)

Key features and details

  • Mouse monoclonal [4A7] to EpCAM - C-terminal
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG1

You may also be interested in

Secondary
Product image
Goat Anti-Mouse IgG H&L (HRP) (ab205719)
Knockout
Product image
Human EPCAM knockout A-431 cell line (ab261902)

View more associated products

Overview

  • Product name

    Anti-EpCAM antibody [4A7] - C-terminal
    See all EpCAM primary antibodies
  • Description

    Mouse monoclonal [4A7] to EpCAM - C-terminal
  • Host species

    Mouse
  • Tested applications

    Suitable for: WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide corresponding to Human EpCAM. (From intracellular domain of the C-terminus).
    Database link: P16422

  • Positive control

    • WB: Wild-type HAP1, MCF7, and HT-29 cell lysates. IHC-P: Human colon adenocarcinoma tissue.
  • General notes

    This product is FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

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 long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.60
    Preservative: 0.1% Sodium azide
    Constituents: PBS, 1% BSA
  • Concentration information loading...
  • Purity

    Protein A/G purified
  • Clonality

    Monoclonal
  • Clone number

    4A7
  • Isotype

    IgG1
  • Research areas

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

Associated products

  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
  • KO cell lines

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

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

    • HT-29 whole cell lysate (ab3952)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab224826 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
1/400. Predicted molecular weight: 35 kDa.

Primary incubation for 1 hour at room temperature.

IHC-P
1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Primary incubation for 10 minutes at room temperature.

Notes
WB
1/400. Predicted molecular weight: 35 kDa.

Primary incubation for 1 hour at room temperature.

IHC-P
1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Primary incubation for 10 minutes at room temperature.

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
    • Omim: 185535 Human
    • SwissProt: P16422 Human
    • Unigene: 542050 Human
    • 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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EpCAM antibody [4A7] - C-terminal (ab224826)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EpCAM antibody [4A7] - C-terminal (ab224826)

      Formalin-fixed, paraffin-embedded human colon adenocarcinoma tissue stained for EpCAM using ab224826 at 1/100 dilution in immunohistochemical analysis.

    • Western blot - Anti-EpCAM antibody [4A7] - C-terminal (ab224826)
      Western blot - Anti-EpCAM antibody [4A7] - C-terminal (ab224826)
      All lanes : Anti-EpCAM antibody [4A7] - C-terminal (ab224826) at 1/400 dilution

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

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 35 kDa
      Observed band size: 35 kDa



      Lanes 1 - 4: Merged signal (red and green). Green - ab224826 observed at 35 kDa. Red - loading control, ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55kDa.

      ab224826 was shown to react with EpCAM in A431 wild-type cells in Western blot. Loss of signal was observed when EpCAM knockout sample was used. A431 wild-type and EpCAM knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% Milk in TBS-T (0.1% Tween®) before incubation with ab224826 and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4°C at a 1 in 400 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-EpCAM antibody [4A7] - C-terminal (ab224826)
      Western blot - Anti-EpCAM antibody [4A7] - C-terminal (ab224826)
      Anti-EpCAM antibody [4A7] - C-terminal (ab224826) at 1/100 dilution + HT-29 (human colorectal adenocarcinoma cell line) cell lysate

      Predicted band size: 35 kDa

    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 (1)

    Publishing research using ab224826? Please let us know so that we can cite the reference in this datasheet.

    ab224826 has been referenced in 1 publication.

    • Keren L  et al. MIBI-TOF: A multiplexed imaging platform relates cellular phenotypes and tissue structure. Sci Adv 5:eaax5851 (2019). Mass Cytometry . PubMed: 31633026

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