United Kingdom (UK)
Recombinant monoclonal antibody to EpCAM. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma HEA125.
UniProt Accession Number of Target Protein: P16422
Alternative Name(s) of Target: CD326; Ep-CAM; Epithelial cell adhesion molecule; Adenocarcinoma-associated antigen; Cell surface glycoprotein Trop-1; Epithelial cell surface antigen; Epithelial glycoprotein; EGP; Epithelial glycoprotein 314; EGP314; hEGP314; KS 1/4 antigen; KSA; Major gastrointestinal tumor-associated protein GA733-2; Tumor-associated calcium signal transducer 1; HEA 125
Immunogen: This antibody was raised by immunising BALB/c mice with the human colon carcinoma cell line HT-29.
Specificity: This antibody is specific for the extracytoplasmic part of human EpCAM, a stable and widely-distributed marker expressed on the surface of many adenocarcinomas. It reacts with carcinomas derived from the colorectum, stomach, pancreas, liver, lung, mammary gland, ovary, thyroid, kidney, urinary bladder and prostate.
Application Notes: This antibody has applications in the detection of normal and neoplastic epithelial cells, and distinguishing carcinomas from non-epithelial neoplasms (Momburg et al, 1987). This antibody has been used in radioimmunoassays on live cells and immunoenzymatic staining assays on glutaraldehyde-fixed cells, as well as in immunoperoxidase staining of frozen and formalin-fixed paraffin tissue sections (Moldenhauer et al, 1987; Momburg et al, 1987). These revealed it to react with most normal human epithelial cells, but not non-epithelial tissue, and to intensely react with human carcinomas and metastatic lesions, but not lymphoma, melanoma, sarcoma or neuroblastoma cell lines (Moldenhauer et al, 1987; Momburg et al, 1987). Squamous cell carcinomas stain less strongly than adenocarcinomas, and keratinizing areas of tumor masses remain negative (Momburg et al, 1987). This antibody has also been used in immunoprecipitation analysis to characterise human EpCAM (Moldenhauer et al, 1987), and in immunohistochemical analysis of EpCAM expression in formalin-fixed, paraffin-embedded liver tissue sections (Igarashi et al, 2013; Wang et al, 2018) and frozen liver tissue sections (Crosby et al, 1998). Additionally, it has been used in the immunocytochemical detection of EpCAM expression in mononuclear cell preparations obtained from human peripheral blood or lamina propria (Qiao et al, 1996).
Antibody first published in:
Moldenhauer et al. Epithelium-specific surface glycoprotein of Mr 34,000 is a widely distributed human carcinoma marker. Br J Cancer. 1987 Dec; 56(6): 714–721. PMID:2449234
Note on publication:
Describes the original generation of this antibody, and its characterisation.
Western Blot using anti-EpCAM antibody HEA125 (Ab01421). human breast carcinoma(A) and human colorectal carcinoma(B) tissue lysate (35µg protein in RIPA buffer) was resolved on an SDS PAGE gel and blots probed with the chimeric mouse IgG version of HEA125 (Ab01421-1.1) at 1 µg/ml before detection using an anti-mouse secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence.
Immunofluorescence staining of Caco-2 cells using anti-EpCAM (Ab01421) HEA125 Immunofluorescence analysis of paraformaldehyde fixed Caco-2 cells stained with the chimeric mouse IgG version of HEA125 (Ab01421-1.1) at 10 µg/ml followed by Alexa Fluor® 488 secondary antibody (2 µg/ml), showing membrane staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom Ab01215-1.1, DAPI, merged channels and an isotype control. The isotype control was stained with anti-unknown antibody (Ab00178-1.1) followed by Alexa Fluor® 488 secondary antibody.
Flow-cytometry using the anti-EpCAM antibody HEA125 (Ab01421). Caco-2 cells were stained with anti-unknown IgG antibody (Ab00178-1.1; isotype control, black line) or the mouse IgG-chimeric version of HEA125 (Ab01421-1.1, blue line) at a dilution of 1:100 for 1h at RT. After washing, bound antibody was detected using a goat anti-mouse IgG AlexaFluor® 488 antibody at a dilution of 1:1000 and cells analyzed using a FACSCanto flow-cytometer.
Western Blot using anti-EpCAM antibody HEA125 (Ab01421). human breast carcinoma(A) and human colorectal carcinoma(B) tissue lysate (35µg protein in RIPA buffer) was resolved on an SDS PAGE gel and blots probed with the chimeric mouse IgG version of HEA125 (Ab01421-1.1) at 1 µg/ml before detection using an anti-mouse secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence.
Flow cytometry using the Anti-EpCAM antibody HEA125 (Ab01421). Caco-2 cells were stained with anti-unknown specificity antibody (Ab00178-1.1; isotype control, black line) or the mouse IgG1 version of HEA125 (Ab01421-1.1, blue line) at a dilution of 1:100 for 1h at RT. After washing, the bound antibody was detected using a goat anti-mouse IgG AlexaFluor® 488 antibody at a dilution of 1:1000 and cells analyzed using a FACSCanto flow-cytometer.