PathScan^® EMT Duplex IF Kit #7771

Products Included	No.	Volume	Applicaton	Dilution	Reactivity	Homology†
PathScan® EMT Duplex IF Kit Primary Antibody Cocktail	7783	100 µl	Immunofluorescence (Immunocytochemistry), Immunofluorescence (Paraffin)	1:100	Human	Mouse
PathScan® Duplex IF Kit Detection Cocktail I	7832	100 µl	Immunofluorescence (Immunocytochemistry), Immunofluorescence (Paraffin)	1:100	N/A	N/A

†Species predicted to react based on 100% sequence homology.

Kit Analytes	Detection Dye	Ex(max) (nm)	Em(max) (nm)
E-cadherin	Alexa Fluor® 488	495	519
Vimentin	Alexa Fluor® 555	555	565

The PathScan^® EMT Duplex IF Kit offers a novel method to simultaneously monitor cells of epithelial or mesenchymal origin, as well as those undergoing an epithelial-mesenchymal transition (EMT) using manual immunofluorescence microscopy or automated imaging and laser scanning high content platforms. This kit contains a cocktail of two high quality primary antibodies targeted against vimentin and E-cadherin, as well as a detection cocktail utilizing the Alexa Fluor^® series of fluorescent dyes. Antibody and dye pairings have been pre-optimized and each kit contains enough reagents for 100 assays (based on a working volume of 100 μl/test).

Vimentin mAb detects endogenous levels of total vimentin protein. E-cadherin mAb detects endogenous levels of total E-cadherin protein and does not cross react with related family members such as N-cadherin.

Monoclonal antibodies were produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Arg45 of human vimentin protein or residues surrounding Pro780 of human E-cadherin protein.

Epithelial-mesenchymal transition (EMT) refers to a biological process in which cells undergo a series of biochemical changes that induce a morphological transformation from an epithelial, polarized, adhesive state to an irregular, elongated, mesenchymal phenotype that enables migratory capacity (1,2). EMTs are classified into three subtypes: those involved in implantation, embryogenesis, and organ development; those associated with inflammation and fibrosis; and those involved in invasion and metastasis (1). Molecular changes that are associated with cells during this transformation include the loss of E-cadherin and gain of vimentin expression, hallmark epithelial and mesenchymal markers, respectively (3-5). Numerous studies have established that EMT is an essential step in cancer metastasis (5-7). E-cadherin is regarded as an active suppressor of invasion and tumorigenesis (8). In response to extracellular stimuli, vimentin coordinates various signaling pathways to induce spatial reorganization and structural changes (9), reminiscent of the EMT phenotype observed in motile cells involved in invasion and metastasis (6).

1. Kalluri, R. and Weinberg, R.A. (2009) J Clin Invest 119, 1420-8.
2. Wheelock, M.J. and Johnson, K.R. (2003) Annu Rev Cell Dev Biol 19, 207-35.
3. Lee, J.M. et al. (2006) J Cell Biol 172, 973-81.
4. Yan, W. et al. (2010) J Biol Chem 285, 14042-51.
5. Sethi, S. et al. (2011) Transl Oncol 4, 222-6.
6. Helfand, B.T. et al. (2004) J Cell Sci 117, 133-41.
7. Kim, J.H. et al. (2007) J Korean Med Sci 22, 898-904.
8. Emadi Baygi, M. et al. (2010) Cell Biol Toxicol 26, 553-67.
9. Sethi, S. et al. (2010) Am J Transl Res 3, 90-9.

• Data sheet (168KB)
• MSDS PDF