PathScan® EMT Duplex IF Kit (#7771) Datasheet with Images Cell Signaling Technology

For Research Use Only. Not for Use in Diagnostic Procedures.

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

Applications:

IF-P, IF-IC

UniProt ID:

#P08670, #P12830

Entrez-Gene Id:

7431, 999

Product Information

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

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.

Species Reactivity:

Human

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.

Product Description

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).

Background

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).

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Applications Key

IF-P: Immunofluorescence (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

PathScan is a trademark of Cell Signaling Technology, Inc.

DRAQ5 is a registered trademark of Biostatus Limited.

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 [email protected]. Alexa Fluor® is a registered trademark of Molecular Probes, Inc.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

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