CellSimple™ Epithelial to Mesenchymal Transition Antibody Assay Kit is a fluorescent assay designed for use with the CellSimple™ Cell Analyzer. The kit includes E-Cadherin (24E10) Rabbit mAb (Alexa Fluor® 488) #3199 and Vimentin (D21H3) XP® Rabbit mAb (PE Conjugate) #12020, that can differentiate between epithelial and mesenchymal cell origins, respectively. Thus mean fluorescence intensity (MFI) in the green (525/45 nm) and red (561 nm LP) channels can be used as an indicator for cells undergoing epithelial-mesenchymal transition (EMT).
The conjugated antibodies in this kit are supplied in PBS (pH 7.2), less than 0.1% sodium azide and 2 mg/ml BSA. Store at 4°C. Do not aliquot the antibodies. Protect from light. Do not freeze. The remaining components of the kit should be stored at the recommended temperature upon receipt. All components in this kit are stable for at least 12 months when stored at the recommended temperature and left unused.
Antibodies provided in the CellSimple™ Epithelial to Mesenchymal Transition Antibody Assay Kit detect endogenous levels of their respective target.
The CellSimple™ Cell Analyzer is a benchtop instrument that utilizes a disposable thin-film cassette and a combination of a 488 nm laser, two photomultiplier tubes (525/45 nm and 561 nm LP filters), Coulter Principle-based cell measurements, and on-board software to provide easy-to-run applications and data analysis. Data acquisition occurs within approximately 10 seconds per test. The instrument relies on disposable cassettes for sample handling, which alleviates the need for flow cell cleaning and fluidics maintenance and the instrument is small enough to be portable between the lab bench and the hood. Applications include quantitative assessments of cell viability, apoptosis, other labeled antibody markers and single and multiplexed bead-based assays for protein and cellular analysis.
Epithelial-mesenchymal transition (EMT) is an essential process during development whereby epithelial cells aquire mesenchymal, fribroblast-like properties and display reduced intracellular adhesion and increased motility. This is a critical feature of normal embryonic development, which is also utilized by malignant epithelial tumors to spread beyond their origin (1-3). This tightly regulated process is associated with a number of cellular and molecular events. EMT depends on a reduction in expression of cell adhesion molecules. Cadherins mediate calcium-dependent cell-cell adhesion and play critical roles in normal tissue development (4). E-cadherin is considered to be an active suppressor of invasion and growth in many epithelial cancers (4-6). Recent studies indicate that cancer cells have upregulated N-cadherin in addition to loss of E-cadherin. This change in cadherin expression is called the "cadherin switch" and downregulation of E-cadherin is one of the hallmarks of EMT (1). Cells undergoing EMT may also continue to express epithelial markers and show epithelial morphology while exhibiting increased expression of mesenchymal markers such as vimentin (7).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. CellSimple is a trademark of Cell Signaling Technology, Inc. XP is a registered trademark of Cell Signaling Technology, Inc. The Alexa Fluor dye conjugates in this product are sold under license from Life Technologies Corporation, for research use only excluding use in combination with DNA microarrays and high content screening (HCS).