Product Pathways - Development
PathScan® EMT Duplex IF Kit #7771
|Products Included||Product No.||Volume||Applicaton||Dilution||Reactivity|
|PathScan® EMT Duplex IF Kit Primary Antibody Cocktail||7783||100 µl||IF-IC, IF-P*||1:100||H, (M)|
|PathScan® Duplex IF Kit Detection Cocktail I||7832||100 µl||IF-IC, IF-P*||1:100||n/a|
|Kit Analytes||Detection Dye||Ex(max) (nm)||Em(max) (nm)|
|E-cadherin||Alexa Fluor® 488||495||519|
|Vimentin||Alexa Fluor® 555||555||565|
Reactivity Key: H=Human
Species enclosed in parentheses are predicted to react based on 100% sequence homology.
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).
Specificity / Sensitivity
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.
Confocal immunofluorescent analysis of cocultured MCF7 and MDA-MB-231 cells using PathScan® EMT Duplex IF Kit. Green = E-cadherin, red = vimentin, and blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Source / Purification
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).
- Kalluri, R. and Weinberg, R.A. (2009) J Clin Invest 119, 1420-8.
- Lee, J.M. et al. (2006) J Cell Biol 172, 973-81.
- Yan, W. et al. (2010) J Biol Chem 285, 14042-51.
- Sethi, S. et al. (2011) Transl Oncol 4, 222-6.
- Kim, J.H. et al. (2007) J Korean Med Sci 22, 898-904.
- Emadi Baygi, M. et al. (2010) Cell Biol Toxicol 26, 553-67.
- Sethi, S. et al. (2010) Am J Transl Res 3, 90-9.
- Wheelock, M.J. and Johnson, K.R. (2003) Annu Rev Cell Dev Biol 19, 207-35.
- Helfand, B.T. et al. (2004) J Cell Sci 117, 133-41.
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* Product-specific protocol.
The transfer of the secondary cocktail contained in this kit is contingent on the buyer using the purchased product solely in research conducted by the buyer (whether the buyer is an academic or for-profit entity), for immunocytochemistry, high content screening (HCS) analysis, or flow cytometry applications. The sale of this product is expressly conditioned on the buyer not using the product or its components (1) in manufacturing; (2) to provide a service, information, or data to an unaffiliated third party for payment; (3) for therapeutic, diagnostic or prophylactic purposes; (4) resale, whether or not such product or its components are resold for use in research; or for any other commercial purpose. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@lifetech.
com. Alexa Fluor® is a registered trademark of Molecular Probes, Inc.
For Research Use Only. Not For Use In Diagnostic Procedures.