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EpCAM Recombinant Rabbit Monoclonal Antibody [PS01-69]

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Specification

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Catalog# HA721644

EpCAM Recombinant Rabbit Monoclonal Antibody [PS01-69]

Application

  • IHC-P

  • IF-Cell

  • FC

Reactivity

  • Human

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

EpCAM Recombinant Rabbit Monoclonal Antibody [PS01-69]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human EpCAM aa1-295.

Species Reactivity

Human

Validated Applications

IHC-P, IF-Cell, FC

Target Molecular Weight

Predicted band size: 35 kDa

Positive Control

Human colon tissue, HT-29.

Conjugation

unconjugated

Clone Number

PS01-69

RRID

AB_3072757

Product Features

Form

Liquid

Concentration

1 mg/mL.(The concentration of this product may be batch-dependent)

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • IHC-P

  • 1:200

  • IF-Cell

  • 1:500

  • FC

  • 1:500-1:1,000

Target

Function

EPCAM is a carcinoma-associated antigen and belongs to a family which includes at least 2 type I membrane proteins. The EPCAM protein has a role in embryonic stem cells proliferation and differentiation. EPCAM is used as a target for immunotherapy treatment of human carcinomas. EPCAM is expressed on most normal epithelial cells and gastrointestinal carcinomas and acts as a homotypic calcium-independent cell adhesion molecule. Epithelial cell adhesion molecules (EPCAM) can act as a physical homophilic interaction molecule between intestinal epithelial cells (IECs) and intraepithelial lymphocytes (IELs) at the mucosal epithelium for supplying immunological barrier as a first line of defense against mucosal infection. EPCAM gene mutations result in congenital tufting enteropathy.

Background References

1. Li D et al. EpCAM-targeting CAR-T cell immunotherapy is safe and efficacious for epithelial tumors. Sci Adv. 2023 Dec

2. Li F et al. EpCAM-Targeting Aptamer Radiotracer for Tumor-Specific PET Imaging. Bioconjug Chem. 2021 Jun

Sequence Similarity

Belongs to the EPCAM family.

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.

Post-translational Modification

Hyperglycosylated in carcinoma tissue as compared with autologous normal epithelia. Glycosylation at Asn-198 is crucial for protein stability.

Subcellular Location

Lateral cell membrane, Cell junction.

UNIPROT

SwissProt: P16422 Human

Synonyms

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

Expand

Images

  • Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-EpCAM antibody (<a href="/products/HA721644" style="font-weight: bold;text-decoration: underline;">HA721644</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA721644" style="font-weight: bold;text-decoration: underline;">HA721644</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-EpCAM antibody (HA721644) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721644) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Application: Immunocytochemistry (IF-cell)<br /><br />Species: Human<br />Sample: HT-29 (Human colorectal adenocarcinoma cell) / NCI-H1299 (Human non-small cell Lung carcinoma cell)<br /><br />Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.<br />Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.<br />Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.<br />Antibody dilution buffer: 1% BSA in PBST.<br /><br />Primary antibody: <a href="/products/HA721644" style="font-weight: bold;text-decoration: underline;">HA721644</a>, 1/500, overnight at 4℃.<br />Secondary antibody: Goat Anti-Rabbit IgG (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>), 45 minutes at room temperature.<br />Counterstain: Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).<br /><br />NCI-H1299 is a low EpCAM-expressing cell line (PMID: 39105395).

    ☑ Relative expression (RE)

    Application: Immunocytochemistry (IF-cell)

    Species: Human
    Sample: HT-29 (Human colorectal adenocarcinoma cell) / NCI-H1299 (Human non-small cell Lung carcinoma cell)

    Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
    Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
    Antibody dilution buffer: 1% BSA in PBST.

    Primary antibody: HA721644, 1/500, overnight at 4℃.
    Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
    Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).

    NCI-H1299 is a low EpCAM-expressing cell line (PMID: 39105395).

  • Flow cytometric analysis of HT-29 cells labeling EpCAM.<br /><br />Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (<a href="/products/HA721644" style="font-weight: bold;text-decoration: underline;">HA721644</a>, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of HT-29 cells labeling EpCAM.

    Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (HA721644, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

  • Application: Immunocytochemistry (IF-cell)<br /><br />Species: Human<br />Sample: MCF7 (Human breast cancer cell)<br /><br />Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.<br />Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.<br />Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.<br />Antibody dilution buffer: 1% BSA in PBST.<br /><br />Primary antibody: <a href="/products/HA721644" style="font-weight: bold;text-decoration: underline;">HA721644</a>, 1/500, overnight at 4℃.<br />Secondary antibody: Goat Anti-Rabbit IgG (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>), 45 minutes at room temperature.<br />Counterstain: Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).

    Application: Immunocytochemistry (IF-cell)

    Species: Human
    Sample: MCF7 (Human breast cancer cell)

    Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
    Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
    Antibody dilution buffer: 1% BSA in PBST.

    Primary antibody: HA721644, 1/500, overnight at 4℃.
    Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
    Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).

  • Application: Immunocytochemistry (IF-cell)<br /><br />Species: Human<br />Sample: HCT 116 (Human colon cancer cell)<br /><br />Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.<br />Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.<br />Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.<br />Antibody dilution buffer: 1% BSA in PBST.<br /><br />Primary antibody: <a href="/products/HA721644" style="font-weight: bold;text-decoration: underline;">HA721644</a>, 1/500, overnight at 4℃.<br />Secondary antibody: Goat Anti-Rabbit IgG (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>), 45 minutes at room temperature.<br />Counterstain: Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).

    Application: Immunocytochemistry (IF-cell)

    Species: Human
    Sample: HCT 116 (Human colon cancer cell)

    Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
    Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
    Antibody dilution buffer: 1% BSA in PBST.

    Primary antibody: HA721644, 1/500, overnight at 4℃.
    Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
    Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).

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

Citation

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