EP-CAM Monoclonal Antibody (ZM131), MonoMab™

Product Specific Information

This product is diluted and in a ready-to-use formulation.

A recommended positive control tissue for this product is Colon adenocarcinoma, however positive controls are not limited to this tissue type.

The primary antibody is intended for laboratory professional use in the detection of the corresponding protein in formalin-fixed, paraffin-embedded tissue stained in manual qualitative immunohistochemistry (IHC) testing. This antibody is intended to be used after the primary diagnosis of tumor has been made by conventional histopathology using non-immunological histochemical stains.

EGP40 is a 40-43 kDa transmembrane epithelial glycoprotein, also identified as epithelial specific antigen (ESA), or epithelial cellular adhesion molecule (Ep-CAM). It is expressed on basolateral cell surface in most simple epithelia and a vast majority of carcinomas. This antibody has been used to distinguish adenocarcinoma from pleural mesothelioma and hepatocellular carcinoma. This antibody is also useful in distinguishing serous carcinomas of the ovary from mesothelioma.

Antibody is used with formalin-fixed and paraffin-embedded sections. Pretreatment of deparaffinized tissue with heat-induced epitope retrieval or enzymatic retrieval is recommended. In general, immunohistochemical (IHC) staining techniques allow for the visualization of antigens via the sequential application of a specific antibody to the antigen (primary antibody), a secondary antibody to the primary antibody (link antibody), an enzyme complex and a chromogenic substrate with interposed washing steps. The enzymatic activation of the chromogen results in a visible reaction product at the antigen site. Results are interpreted using a light microscope and aid in the differential diagnosis of pathophysiological processes, which may or may not be associated with a particular antigen.

A positive tissue control must be run with every staining procedure performed. This tissue may contain both positive and negative staining cells or tissue components and serve as both the positive and negative control tissue. External Positive control materials should be fresh autopsy/biopsy/surgical specimens fixed, processed and embedded as soon as possible in the same manner as the patient sample (s). Positive tissue controls are indicative of correctly prepared tissues and proper staining methods. The tissues used for the external positive control materials should be selected from the patient specimens with well-characterized low levels of the positive target activity that gives weak positive staining. The low level of positivity for external positive controls is designed to ensure detection of subtle changes in the primary antibody sensitivity from instability or problems with the staining methodology. A tissue with weak positive staining is more suitable for optimal quality control and for detecting minor levels of reagent degradation.

Internal or external negative control tissue may be used depending on the guidelines and policies that govern the organization to which the end user belongs to. The variety of cell types present in many tissue sections offers internal negative control sites, but this should be verified by the user. The components that do not stain should demonstrate the absence of specific staining, and provide an indication of non-specific background staining. If specific staining occurs in the negative tissue control sites, results with the patient specimens must be considered invalid.

Target Information

Ep-CAM (epithelial adhesion molecule, epithelial specific antigen, ESA) is a transmembrane glycoprotein expressed in the epithelium with a molecular weight of approximately 40 kDa, which functions as an epithelial cell adhesion molecule. Ep-CAM functions as a homotypic calcium-independent cell adhesion molecule, and has a direct impact on cell cycle, proliferation and metabolism of epithelial cells and fibroblasts due to its ability to rapidly induce the proto-oncogene c-myc and the cell cycle regulating genes cyclin A and E. Ep-CAM mediates Ca2+-independent homotypic interactions. Formation of Ep-CAM-mediated adhesions have a negative regulatory effect on adhesions mediated by classic cadherins, which may have strong effects on the differentiation and growth of epithelial cells. Ep-CAM overexpression was suggested to be associated with enhanced epithelial proliferation. Ep-CAM is highly expressed in human carcinomas, and is a marker for tumors of epithelial lineage. Ep-CAM is expressed on baso-lateral cell surface in most simple epithelia and many carcinoma types. Also, Ep-CAM reportedly distinguishes adenocarcinomas from pleural mesotheliomas.