Epithelial-Mesenchymal Transition (EMT)

1 Store at -20 C. Epithelial-Mesenchymal Transition (EMT). Antibody Sampler Kit 3 1 Kit n Orders n 877-616-CELL (2355). (9 x 20 l) #9782. Support n 877-678-TECH (8324). Web n rev. 12/04/17. For Research Use Only. Not For Use In Diagnostic Procedures. Storage: Supplied in 10 mM sodium HEPES (pH ), 150. Products Included Product # Quantity Mol. Wt. Isotype mM NaCl, 100 g/ml BSA, 50% glycerol and less than Vimentin (D21H3) XP Rabbit mAb 5741 20 l 57 kDa Rabbit IgG sodium azide. Store at 20 C. Do not aliquot the antibodies. N-Cadherin (D4R1H) XP Rabbit mAb 13116 20 l 140 kDa Rabbit IgG Recommended Antibody Dilutions: Western blotting 1:1000.

2 Claudin-1 (D5H1D) XP Rabbit mAb . 13255 20 l 20 kDa Rabbit IgG. Please visit for validation data -Catenin (D10A8) XP Rabbit mAb 8480 20 l 92 kDa Rabbit IgG and a complete listing of recommended companion ZO-1 (D7D12) Rabbit mAb 8193 20 l 220 kDa Rabbit IgG products. Snail (C15D3) Rabbit mAb 3879 20 l 29 kDa Rabbit IgG. Slug (C19G7) Rabbit mAb 9585 20 l 30 kDa Rabbit IgG. TCF8/ZEB1 (D80D3) Rabbit mAb 3396 20 l 200 kDa Rabbit IgG. E-Cadherin (24E10) Rabbit mAb 3195 20 l 135 kDa Rabbit IgG. Anti-rabbit IgG, HRP-linked Antibody 7074 100 l Goat See for individual component applications, species cross-reactivity, dilutions, and additional application protocols.

3 Description: The Epithelial-Mesenchymal Transition (EMT) eton (7,8). Zona occludens proteins ZO-1, 2, and 3 (also recognizes endogenous levels of total N-cadherin protein. Antibody Sampler Kit provides an economical means known as TJP 1, 2, and 3) are peripheral membrane adaptor Claudin-1 (D5H1D) XP Rabbit mAb recognizes endog- of evaluating EMT. The kit includes enough antibody to proteins that link junctional transmembrane proteins such enous levels of total claudin-1 protein. ZO-1 (D7D12) Rabbit perform two western blot experiments with each per primary as occludin and claudin to the actin cytoskeleton (9). ZO-1 mAb detects endogenous levels of total ZO-1 protein.

4 Antibody. and -2 are required for tight junction formation and function Vimentin (D21H3) XP Rabbit mAb detects endogenous (10,11); mutations in ZO-1 and Claudin induce EMT (12). levels of total vimentin protein. Snail (C15D3) Rabbit mAb Background: Epithelial-Mesenchymal Transition (EMT) is Vimentin is an intermediate filament of mesenchymal origin detects endogenous levels of total Snail protein. Slug an essential process during development whereby epithelial and is present at early developmental stages. Vimentin's (C19G7) Rabbit mAb detects endogenous levels of total cells aquire mesenchymal , fibroblast-like properties and dynamic structural changes and spatial re-organization in Slug protein.

5 TCF8/ZEB1 (D80D3) Rabbit mAb detects display reduced intracellular adhesion and increased motility. response to extracellular stimuli helps to coordinate various endogenous levels of total TCF8/ZEB1 protein. -Catenin This is a critical feature of normal embryonic development, signaling pathways (13). -catenin is a key downstream (D10A8) XP Rabbit mAb detects endogenous levels of which is also utilized by malignant epithelial tumors to XP and Cell Signaling Technology are trademarks of Cell Signaling Technology, Inc. effector in the Wnt signaling pathway (14). It is implicated in total -catenin protein. spread beyond their origin (1-3).

6 This tightly regulated two major biological processes in vertebrates: early embry- process is associated with a number of cellular and molecular Source/Purification: Monoclonal antibodies are onic development (15) and tumorigenesis (16). -catenin events. EMT depends on a reduction in expression of cell produced by immunizing animals with a synthetic peptide also activates Slug. Slug (SNAI2) is a widely expressed adhesion molecules. corresponding to residues surrounding Arg526 of human transcriptional repressor and member of the Snail family of Cadherins mediate calcium-dependent cell-cell adhesion N-cadherin protein, a synthetic peptide corresponding to zinc finger transcription factors (17).

7 Similar to the related and play critical roles in normal tissue development (4). residues near the carboxy terminus of human claudin-1. Snail protein, Slug binds to the E-cadherin promoter region E-cadherin is considered an active suppressor of invasion protein, a synthetic peptide corresponding to residues near to repress transcription during development (18). The binding and growth of many epithelial cancers (4-6). Recent studies the carboxy terminus of human ZO-1 protein, a recombinant of Slug to integrin promoter sequences represses integrin indicate that cancer cells have up-regulated N-cadherin human Snail protein, a recombinant human Slug protein, expression and results in reduced cell adhesion (19).

8 Down in addition to loss of E-cadherin. This change in cadherin a synthetic peptide corresponding to residues surrounding regulation of E-cadherin expression occurs during the EMT. expression is called the cadherin switch and downregula- Arg45 of human vimentin protein, a synthetic peptide during embryonic development (20). ZEB family proteins are tion of E-cadherin is one of the hallmarks of EMT (1). Tight corresponding to residues surrounding Pro780 of human zinc finger and homeobox domain containing transcription junctions, or zonula occludens, form a continuous barrier to E-cadherin, a synthetic peptide corresponding to residues 2014 Cell Signaling Technology, Inc.

9 Factors. One of the targets suppressed by ZEB proteins is fluids across the epithelium and endothelium. They function surrounding Asp868 of human TCF8/ZEB1 protein, or a E-cadherin (1). in regulation of paracellular permeability and in the mainte- synthetic peptide corresponding to residues surrounding nance of cell polarity, blocking the movement of transmem- Specificity/Sensitivity: E-Cadherin (24E10) Rabbit mAb Pro714 of human -catenin protein. brane proteins between the apical and the basolateral cell detects endogenous levels of total E-cadherin protein. The surfaces. Tight junctions are composed of claudin and antibody does not cross-react with related family members, occludin proteins, which join the junctions to the cytoskel- such as N-cadherin.

10 N-Cadherin (D4R1H) XP Rabbit mAb Patent No. 5,675,063. Tween 20 is a registered trademark of ICI Americas, Inc. Applications Key: W Western IP Immunoprecipitation IHC Immunohistochemistry ChIP Chromatin Immunoprecipitation IF Immunofluorescence F Flow cytometry E-P ELISA-Peptide Species Cross-Reactivity Key: H human M mouse R rat Hm hamster Mk monkey 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 All all species expected Species enclosed in parentheses are predicted to react based on 100% homology. #9782. Western Immunoblotting Protocol For western blots, incubate membrane with diluted primary antibody in either 5% w/v BSA or nonfat dry milk, 1X TBS, Tween 20 at 4 C with gentle shaking, overnight.