Epithelial-to-Mesenchymal Transition Markers

The Epithelial-to-Mesenchymal Transition (EMT) is a process that normally occurs during embryogenesis, but is also present in wound healing and cancer progression of epithelial tumors. 

Download article

 

The EMT marker antibody panel consists of PrecisA Monoclonals™ targeting key EMT markers for cell junctions, cytoskeletal changes, transcription regulation and migration/motility.

Antibody panel for Epithelial-to-Mesenchymal transition markers

The antibodies targeting selected EMT markers are carefully developed and characterized to give precise and versatile qualities:

  • IHC-validated in relevant normal and cancerous human tissues
  • WB-validated in positive and negative cell lines
  • Available with different isotypes, allowing for multiplexing experiments
  • Supplemented with information on antigens used for immunization and precise epitope sequence

The monoclonal antibodies within the panel have been developed using the same stringent conditions as for all PrecisA Monoclonals, ensuring a secured continuity and stable supply. 

Explore EMT Markers

Explore EMT Markers

The complete EMT marker antibody panel is presented in our product catalog. 

Browse antibodies

Application examples of key EMT markers

Among the EMT marker antibodies are the Anti-CDH1 (AMAb90863) and Anti-CDH2 (AMAb91220) antibodies, targeting E-cadherin and N-cadherin respectively. E-cadherin (CDH1) is a calcium-depend­ent transmembrane protein forming adherence junctions between the epithelial cells. E-cadherin suppresses tumor invasion and metastasis, while down-regulation of E-cadherin expression is a critical molecular feature of EMT. Often, the loss of E-cadherin expression is associated with an increase in N-cadherin (CDH2)  levels, the so-called ‘cadherin switch’. The upregulation of N-cadherin expression in tumor cells induces increased motility, invasion and metastasis.

The Western blot example in the image below shows a band of expected molecular weight in the CDH1-positive MCF7 cell line and the absence of antibody binding in the CDH1-negative SK-BR-3 and HeLa cell lines using the Anti-CDH1 antibody (A). The Anti-CDH2 antibody shows a band of predicted size in the CDH2-positive U-251 MG cell line (B).

Western blot using Anti-CDH1 (AMAb90863) and Anti-CDH2 (AMAb91220) antibodies.
Western blot using Anti-CDH1 (AMAb90863) and Anti-CDH2 (AMAb91220) antibodies. Blot A CDH1: Lane 1: Marker [kDa], Lane 2: Human cell line MCF-7 (positive), Lane 3: Human cell line SK-BR-3 (negative), Lane 4: Human cell line HeLa (negative). Blot B CHD2: Lane 1: Marker [kDa], Lane 2: Human cell line U-251 MG (positive).

As illustrated below, E-cadherin and N-cadherin display a complementary expression pattern in many normal tissues. E-cadherin immunoreactivity is detected in simple and stratified epithelial tissue, e.g. gastrointestinal tract (A) and cervix (B), while it is absent in brain (C), as visualized by IHC staining with the Anti-CDH1 antibody. On the contrary, N-cadherin (CDH2) expression is high in the nervous system (F and inset on D), while glandular epithelium of gastrointestinal tract (D) and squamous epithelium (E) show no protein expression.

E-cadherin (A-C) and N-cadherin (D-F) expression profiles in normal human tissues
E-cadherin (A-C) and N-cadherin (D-F) expression profiles in normal human tissues shown by IHC with the Anti-CDH1 antibody AMAb90863 and the Anti-CDH2 antibody AMAb91220. Note the differential expression of two cadherins in colon (A, D), cervix (B, E) and cerebellum (C, F). Inset on D shows CDH2-immunoreactivity in the peripheral ganglion neurons in rectum.

Transcription factors

Reprogramming of gene expression during EMT occurs in a highly regulated process. IHC images in the image below demonstrate expression of some of the transcriptional factors involved in regulation of the EMT, including SNAI1 (AMAb91215, A)SIX1 (AMAb90544, B) and ZNF703 (AMAb90789, C) in selected cancer tissues.

Transcription factors involved in the regulation of EMT.
Transcription factors involved in the regulation of EMT. IHC images show nuclear immunoreactivity in tumor cells in (A) colorectal cancer (Anti-SNAI1 antibody AMAb91215), (B) cervical cancer (Anti-SIX1 antibody AMAb90544) and (C) breast cancer (Anti-ZNF703 AMAb90789).

Using EMT markers of defined isotypes for multiplexed immunofluorescence

The EMT panel includes monoclonal antibodies with different isotypes, which allows for the simultaneous study of multiple markers using isotype-specific secondary antibodies. 

The images below show multiplexed staining of colorectal cancer tissues derived from two different patients using the LAMC1 (A, E: AMAb91138, IgG2b)Anti-CDH1 (B, F: AMAb90863, IgG1) and Anti-CTNNB1 (C, G: AMAb91209, IgG2a) monoclonal antibodies. These two tumors show different degree of differentiation, as indicated by the preserved basement membrane in Tumor 1 (note the arrows in A), and the absence of basement membrane in Tumor 2 (E). The tumor with a higher degree of differentiation (Tumor 1) shows higher expression of E-cadherin (B) as compared to the tumor with lower differentiation grade (F). Also, note the absence of LAMC1 immunoreactivity in Tumor 2 (E). Beta-catenin (CTNNB1) expression is still pre­served in both tumors (C, G). Panels D and H show overlay images for the two tumors.

Multiplexed IHC-IF staining of two colorectal tumors
Multiplexed IHC-IF staining of two colorectal tumors (A-D and E-H) showing laminin-gamma 1 (A, E), E-cadherin (B, F) and beta-catenin (C, G) immunoreactivity using primary antibodies of different isotypes: Anti-LAMC1 (AMAb91138), IgG2b (blue), Anti-CDH1 AMAb90863, IgG1 (red), and Anti-CTNNB1 AMAb91209, IgG2a (green). Arrows in A indicate basement membrane. Alexa Fluor® 647-, 594- and 488-labelled isotype-specific secondary antibodies (Life Technologies) were used for visualization.