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Metastasis antibodies: choose the most sensitive

Related

  • Cancer biology resources
    • Cell adhesion and metastasis poster
      • Gelatin zymography protocol
        • Collagen protocol tips
          • Choose your MMP9 antibody
            • Measuring activity of gelatinase enzymes
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                    • Immunohistochemistry (IHC) application guide
                      • The complete ELISA guide
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                          ​We compare some of our metastasis antibodies for IHC with the leading alternatives from our competitors. 

                          The IHC images below were made using sections of formalin-fixed paraffin embedded tissue samples. These sections were pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0). All primary antibodies were applied for 15 minutes at room temperature and detected using an HRP-conjugated compact polymer system. DAB was used as the chromogen. The sections were then counterstained with hematoxylin.

                          Results summary

                          • Our N-cadherin, E-cadherin, MMP-2, and collagen I antibodies tested here are all extremely sensitive even at low concentrations
                          • Our antibodies consistently show a specific signal with very low background
                          • Our E-cadherin (ab40772) and MMP-2 (ab86607) antibodies were shown to perform much better than comparable antibodies from leading competitors 

                          ​N-cadherin

                          N-cadherin is well known for its role in promoting epithelial to mesenchymal transition (EMT) during metastasis. During EMT there is an increased expression of N-cadherin leading to a loss of intercellular adhesion and increased cell motility promote tumor cell invasion. Research has also shown that N-cadherin/FGFR plays a role in promoting metastasis through differential regulation of ERK and AKT1. This highlights the potential for targeting the FGFR in certain types of tumors.

                          Figure 1 (below) shows kidney carcinoma stained using our RabMAb® N-cadherin monoclonal antibody (A) compared to a rabbit monoclonal antibody from one of our top competitors. At the same dilution, the RabMAb® N-cadherin monoclonal antibody gives an excellent result with specific staining, strong signal, and low background. Compared to the competitor's antibody it gives a stronger signal. 

                          ​​Figure 1. N-cadherin immunohistochemistry staining in kidney carcinoma. A) Abcam RabMAb® N-cadherin monoclonal antibody (ab76011). B) Competitor N-cadherin rabbit monoclonal antibody. Antibody dilution of 1 µg/mL for A and B. 

                          E-cadherin

                          E-cadherin is also well known for its role in EMT during metastasis. Tumor cells will exhibit a downregulation of E-cadherin on their surface to aid progression through the extracellular matrix. It is known that miR-9, which is upregulated in tumor cells by MYC and MYCN, can directly target E-cadherin mRNA2. Downregulation of E-cadherin by miRNA mediated degradation then leads to increased tumor cell motility and invasiveness. 

                          Figure 2 (below) shows breast carcinoma stained using our RabMAb® E-cadherin monoclonal antibody (A) compared to a rabbit monoclonal antibody from another of our competitors. At a lower dilution, the RabMAb® E-cadherin monoclonal antibody gives much better results compared to the competitor's antibody. The RabMAb® staining is specific with a strong signal and low background. 



                          AB
                          Antibody

                          Dilution

                          Staining description 

                          Verdict

                          ​​Figure 2. E-cadherin immunohistochemistry staining in breast carcinoma. A) Abcam RabMAb® E-cadherin monoclonal antibody (ab40772). B) Competitor E-cadherin rabbit monoclonal antibody. Antibody dilution for A 1/1,000 and B 1/200.

                          MMP-2

                          MMP-2 promotes cancer cell invasion by degrading the ECM allowing tumor cells to migrate to a secondary tumor site during metastasis. Both MMP-2 and MMP-9 will breakdown type IV collagen, the most abundant component found in the basement membrane. It is also known that MMP-2 may activate TGF-β, which will promote EMT, further aiding cancer metastasis3.

                          Figure 3 (below) shows pancreas carcinoma stained using our MMP-2 mouse monoclonal antibody (A) compared to a mouse monoclonal antibody from one of our competitors. At the same dilution, our mouse monoclonal antibody gives an excellent result with specific staining, strong signal, and low background. Compared to the competitor's antibody it gives a much better signal. 

                          ​​Figure 3. MMP-2 immunohistochemistry staining in pancreas carcinoma. A) Abcam MMP-2 mouse monoclonal antibody (ab86607). B) Competitor MMP-2 mouse monoclonal antibody. Antibody dilution of 1 µg/mL for A and B. 

                          Collagen I

                          Collagen I is a key component of the tumor microenvironment. It is found in most connective tissues, tendons, and bone. The ECM undergoes many structural changes during tumor progression, including increased deposition of fibronectin, proteoglycans, and collagens I, III and IV.  Density/stiffness of collagen-dense ECM has been shown to interact with hormonal signals to drive metastasis in some cancer types4. 

                          Figure 4 (below) shows breast carcinoma stained using our collagen I rabbit polyclonal antibody (A) compared to a rabbit polyclonal antibody from our top competitor. At the same dilution, our antibody gives a stronger signal, more specific signal when compared to the competitor's antibody. Although in this case, both antibodies gave excellent results overall. 

                          ​

                          ​Figure 4. Collagen I immunohistochemistry staining in breast carcinoma. A) Abcam collagen I rabbit polyclonal antibody (ab34710). B) Competitor collagen I rabbit polyclonal antibody. Antibody dilution of 5 µg/mL for A and B. 


                          References

                          1) Qian X, Anzovino A, Kim S, Suyama K, Yao J, Hulit J, Agiostratidou G, Chandiramani N, McDaid HM, Nagi C, Cohen HW, Phillips GR, Norton L, Hazan RB. (2014) N-cadherin/FGFR promotes metastasis through epithelial-to-mesenchymal transition and stem/progenitor cell-like properties. Oncogene, 3411-21

                          2) Ma L, Young J, Prabhala H, Pan E, Mestdagh P, Muth D, Teruya-Feldstein J, Reinhardt F, Onder T.T, Valastyan S, Westermann F, Speleman F, Vandesompele J, and Weinberg R.A. (2010) miR-9, a MYC/MYCN-activated microRNA, regulates E-cadherin and cancer metastasis. Nature Cell Biology, 247–256 

                          3) Gialeli C, Theocharis A.D, Karamanos N.K. (2011) Roles of matrix metalloproteinases in cancer progression and their pharmacological targeting. The FEBS Journal, 16–27

                          4) Barcus C, O’Leary K, Brockman J, Rugowski D, Liu Y, Garcia N, Yu M, Keely P, Eliceiri K, Schuler L. (2017) Elevated collagen-I augments tumor progressive signals, intravasation and metastasis of prolactin-induced estrogen receptor alpha positive mammary tumor cells. Breast Cancer Res, 19: 9

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