Cell Signaling Technology

Product Pathways - Cytoskeletal Signaling

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit #9782

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
N-Cadherin Antibody #4061 40 µl W H M R Mk (Dg) 140 Rabbit
Claudin-1 Antibody #4933 40 µl W IP H (M) (R) 20 Rabbit
ZO-1 (D7D12) Rabbit mAb #8193 40 µl W IP H Mk 220 Rabbit IgG
Vimentin (D21H3) XP® Rabbit mAb #5741 40 µl W IHC-P IF-IC F H M R Mk 57 Rabbit IgG
E-Cadherin (24E10) Rabbit mAb #3195 40 µl W IHC-P IHC-F IF-IC F H M (B) (Dg) (Pg) 135 Rabbit IgG
Snail (C15D3) Rabbit mAb #3879 40 µl W IP H M R Mk 29 Rabbit IgG
Slug (C19G7) Rabbit mAb #9585 40 µl W IP IHC-P IF-IC H M 30 Rabbit IgG
TCF8/ZEB1 (D80D3) Rabbit mAb #3396 40 µl W IP H (M) (R) 200 Rabbit IgG
β-Catenin (D10A8) XP® Rabbit mAb #8480 40 µl W IP IHC-P IHC-F IF-F IF-IC F ChIP H M R Mk (Z) (B) (Pg) (GP) (Hr) 92 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl Goat

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IHC-F=Immunohistochemistry (Frozen)  IF-F=Immunofluorescence (Frozen)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry  ChIP=Chromatin IP
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  Z=Zebrafish  B=Bovine  Dg=Dog  Pg=Pig  GP=Guinea Pig  Hr=Horse
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

E-Cadherin (24E10) Rabbit mAb detects endogenous levels of total E-cadherin protein. The antibody does not cross-react with related family members, such as N-cadherin. N-Cadherin Antibody detects endogenous levels of total N-cadherin protein. This antibody does not cross-react with other cadherin family members. Claudin-1 Antibody detects endogenous levels of total claudin-1 protein. Based on sequence similarity, the antibody may cross-react with claudin-2. ZO-1 (D7D12) Rabbit mAb detects endogenous levels of total ZO-1 protein. Vimentin (D21H3) XP® Rabbit mAb detects endogenous levels of total vimentin protein. Snail (C15D3) Rabbit mAb detects endogenous levels of total Snail protein. Slug (C19G7) Rabbit mAb detects endogenous levels of total Slug protein. TCF8/ZEB1 (D80D3) Rabbit mAb detects endogenous levels of total TCF8/ZEB1 protein. β-Catenin (D10A8) XP® Rabbit mAb detects endogenous levels of total β-catenin protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using E-Cadherin (24E10) Rabbit mAb #3195.

Western Blotting

Western Blotting

Western blot analysis of extracts from Jurkat, HT-1080 and A172 cells using TCF8/ZEB1 (D80D3) Rabbit mAb #3396.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Snail (C15D3) Rabbit mAb #3879.


Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using N-Cadherin Antibody #4061.

Western Blotting

Western Blotting

Western blot analysis of extracts from A-431 and ZR-75 cells using Claudin-1 Antibody #4933.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Vimentin (D21H3) XP® Rabbit mAb #5741.


Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using ZO-1 (D7D12) Rabbit mAb #8193.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using β-Catenin (D10A8) XP® Rabbit mAb #8480.

Western Blotting

Western Blotting

Western blot analysis of extracts from A-204, SK-MEL-5, and NIH/3T3 cells using Slug (C19G7) Rabbit mAb #9585.


Description

The Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit provides an economical means of evaluating EMT. The kit contains enough primary antibody to perform four western blots per primary.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with: a synthetic peptide corresponding to residues within the amino terminus of human N-cadherin protein, a synthetic peptide corresponding to the carboxy terminus of mouse claudin-1 protein. Polyclonal antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human ZO-1 protein, a recombinant human Snail protein, a recombinant human Slug protein, a synthetic peptide corresponding to residues surrounding Arg45 of human vimentin protein, a synthetic peptide corresponding to residues surrounding Pro780 of human E-cadherin, a synthetic peptide corresponding to residues surrounding Asp868 of human TCF8/ZEB1 protein, or a synthetic peptide corresponding to residues surrounding Pro714 of human ß-catenin protein.

Background

Epithelial-mesenchymal transition (EMT) is an essential process during development whereby epithelial cells aquire mesenchymal, fibroblast-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 an active suppressor of invasion and growth of many epithelial cancers (4-6). Recent studies indicate that cancer cells have up-regulated 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). Tight junctions, or zonula occludens, form a continuous barrier to fluids across the epithelium and endothelium. They function in regulation of paracellular permeability and in the maintenance of cell polarity, blocking the movement of transmembrane proteins between the apical and the basolateral cell surfaces. Tight junctions are composed of claudin and occludin proteins, which join the junctions to the cytoskeleton (7,8). Zona occludens proteins ZO-1, 2, and 3 (also known as TJP 1, 2, and 3) are peripheral membrane adaptor proteins that link junctional transmembrane proteins such as occludin and claudin to the actin cytoskeleton (9). ZO-1 and -2 are required for tight junction formation and function (10,11); mutations in ZO-1 and Claudin induce EMT (12). Vimentin is an intermediate filament of mesenchymal origin and is present at early developmental stages. Vimentin's dynamic structural changes and spatial re-organization in response to extracellular stimuli helps to coordinate various signaling pathways (13). β-catenin is a key downstream effector in the Wnt signaling pathway (14). It is implicated in two major biological processes in vertebrates: early embryonic development (15) and tumorigenesis (16). β-catenin also activates Slug. Slug (SNAI2) is a widely expressed transcriptional repressor and member of the Snail family of zinc finger transcription factors (17). Similar to the related Snail protein, Slug binds to the E-cadherin promoter region to repress transcription during development (18). The binding of Slug to integrin promoter sequences represses integrin expression and results in reduced cell adhesion (19). Down regulation of E-cadherin expression occurs during the EMT during embryonic development (20). ZEB family proteins are zinc finger and homeobox domain containing transcription factors. One of the targets suppressed by ZEB proteins is E-cadherin (1).

  1. Aigner, K. et al. (2007) Oncogene 26, 6979-88.
  2. Peinado, H. et al. (2007) Nat Rev Cancer 7, 415-28.
  3. Moreno-Bueno, G. et al. (2008) Oncogene 27, 6958-69.
  4. Wheelock, M.J. and Johnson, K.R. (2003) Annu Rev Cell Dev Biol 19, 207-35.
  5. Christofori, G. (2003) EMBO J 22, 2318-23.
  6. Hazan, R.B. et al. (2004) Ann N Y Acad Sci 1014, 155-63.
  7. Shin, K. et al. (2006) Annu Rev Cell Dev Biol 22, 207-35.
  8. Oliveira, S.S. and Morgado-Díaz, J.A. (2007) Cell Mol Life Sci 64, 17-28.
  9. Matter, K. and Balda, M.S. (2007) J Cell Sci 120, 1505-11.
  10. Hernandez, S. et al. (2007) Exp Cell Res 313, 1533-47.
  11. Umeda, K. et al. (2006) Cell 126, 741-54.
  12. Reichert, M. et al. (2000) J Biol Chem 275, 9492-500.
  13. Helfand, B.T. et al. (2004) J Cell Sci 117, 133-41.
  14. Cadigan, K.M. and Nusse, R. (1997) Genes Dev 11, 3286-305.
  15. Wodarz, A. and Nusse, R. (1998) Annu Rev Cell Dev Biol 14, 59-88.
  16. Polakis, P. (1999) Curr Opin Genet Dev 9, 15-21.
  17. Inukai, T. et al. (1999) Mol Cell 4, 343-52.
  18. Bolós, V. et al. (2003) J Cell Sci 116, 499-511.
  19. Turner, F.E. et al. (2006) J Biol Chem 281, 21321-31.
  20. Barrallo-Gimeno, A. and Nieto, M.A. (2005) Development 132, 3151-61.

Application References

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Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc. Selected rabbit monoclonal antibodies produced under license (granting certain rights including those under U. S. Patent No. 5,675,063) from Epitomics, Inc.

For Research Use Only. Not For Use In Diagnostic Procedures.

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