Recombinant
RabMAb

Recombinant Anti-EpCAM antibody [EPR20533-63] (ab221552)

Overview

  • Product name

    Anti-EpCAM antibody [EPR20533-63]
    See all EpCAM primary antibodies
  • Description

    Rabbit monoclonal [EPR20533-63] to EpCAM
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IP, Flow Cyt, ICC/IF, WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse
  • Immunogen

    Recombinant fragment within Mouse EpCAM aa 1-300. The exact sequence is proprietary.
    Database link: Q99JW5

  • Positive control

    • WB: 4T1 whole cell lysate; Mouse colon, kidney and small intestine lysates. IHC-P: Mouse colon and lung tissues. ICC/IF: 4T1 cells. Flow Cyt: 4T1 cells. IP: Mouse kidney lysate; 4T1 whole cell lysate.
  • General notes

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab221552 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
IP 1/30.
Flow Cyt 1/500.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF 1/100.
WB 1/5000. Detects a band of approximately 39, 12 kDa (predicted molecular weight: 35 kDa).
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 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

  • Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling EpCAM with ab221552 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous staining on mouse colon is observed (PMID: 15637741). Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized 4T1 (mouse mammary gland carcinoma cell line) and NIH/3T3 (mouse embyro fibroblast cell line) cells labeling EpCAM with ab221552  at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing Membranous staining on 4T1 cells.

    Negative control: NIH/3T3 (PMID:23264216).

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Immunohistochemical analysis of paraffin-embedded mouse lung tissue labeling EpCAM with ab221552 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous staining on mouse lung is observed (PMID: 15637741). Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

  • All lanes : Anti-EpCAM antibody [EPR20533-63] (ab221552) at 1/5000 dilution

    Lane 1 : 4T1 (mouse mammary gland carcinoma cell line) whole cell lysate
    Lane 2 : NIH/3T3 (mouse embyro fibroblast cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

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

    Developed using the ECL technique.

    Predicted band size: 35 kDa
    Observed band size: 39 kDa
    why is the actual band size different from the predicted?


    Exposure time: 1 second


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The MW observed is consistent with the literature (PMID 23409978; PMID 23618806).

    Negative control: NIH/3T3 (PMID:23264216).

  • All lanes : Anti-EpCAM antibody [EPR20533-63] (ab221552) at 1/5000 dilution

    Lane 1 : Mouse colon lysate
    Lane 2 : Mouse kidney lysate
    Lane 3 : Mouse small intestine lysate

    Lysates/proteins at 20 µg per lane.

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

    Developed using the ECL technique.

    Predicted band size: 35 kDa
    Observed band size: 39 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1/2: 3 seconds; Lane 3: 10 seconds.

    The MW observed is consistent with the literature (PMID 23409978; PMID 23618806).

  • All lanes : Anti-EpCAM antibody [EPR20533-63] (ab221552) at 1/5000 dilution

    Lane 1 : 4T1 (mouse mammary gland carcinoma cell line) whole cell lysate
    Lane 2 : NIH/3T3 (mouse embyro fibroblast cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

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

    Developed using the ECL technique.

    Predicted band size: 35 kDa
    Observed band size: 12,39 kDa why is the actual band size different from the predicted?


    Exposure time: 10 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The MW observed is consistent with the literature (PMID 23618806).

    The band between 12-15kDa has been documented in literature as the cleaved form of EpCAM (PMID: 23409978).

    Negative control: NIH/3T3 (PMID:23264216).

  • Flow cytometric analysis of 4% paraformaldehyde-fixed 4T1 (mouse mammary gland carcinoma cell line) and NIH/3T3 (mouse embyro fibroblast cell line) cells labeling EpCAM with ab221552 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.

    Flow cytometry was performed with fresh cells without fixation and permeabilization.

    Negative control: NIH/3T3 (PMID:23264216).

  • EpCAM was immunoprecipitated from 0.35 mg of mouse kidney lysate with ab221552 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab221552 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: Mouse kidney lysate 10 µg (Input). 

    Lane 2: ab221552 IP in mouse kidney lysate. 

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab221552 in mouse kidney lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 seconds.

  • EpCAM was immunoprecipitated from 0.35 mg of 4T1 (mouse mammary gland carcinoma cell line) whole cell lysate with ab221552 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab221552 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: 4T1 (Mouse mammary gland carcinoma) whole cell lysate 10 µg (Input). 

    Lane 2: ab221552 IP in 4T1 whole cell lysate . 

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab221552 in 4T1 whole cell lysate .

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 30 seconds.

    The band between 12-15kDa has been documented in literature as the cleaved form of EpCAM (PMID: 23409978).

References

ab221552 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

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1-2 of 2 Abreviews

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (gut)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: pH9
Permeabilization
No
Specification
gut
Blocking step
Serum as blocking agent for 10 minute(s) · Concentration: 10% · Temperature: 22°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted May 28 2019

Application
IHC - Wholemount
Sample
Mouse Embryo (E7.5 mouse embryo)
Specification
E7.5 mouse embryo

Abcam user community

Verified customer

Submitted Dec 21 2018

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