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9782
Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit

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

IF-IC Image 1

Confocal immunofluorescent analysis of SNB19 cells using Vimentin (D21H3) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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Western Blotting Image 2

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

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Western Blotting Image 3

Western blot analysis of extracts from A172 and MCF7 cells using N-Cadherin (D4R1H) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

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IHC-P (paraffin) Image 4

Immunohistochemical analysis of paraffin-embedded human colon using N-Cadherin (D4R1H) XP® Rabbit mAb. Note staining of myenteric plexus.

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Western Blotting Image 5

Western blot analysis of extracts from A431 and MCF7 cells using Claudin-1 (D5H1D) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

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Chromatin IP-seq Image 6

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HCT116 cells and either 20 μl of β-Catenin (D10A8) XP® Rabbit mAb or 5 μl of Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb #8814, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared from 5 ng enriched ChIP DNA using NEBNext® Ultra™ II DNA Library Prep Kit for Illumina®, and sequenced on the Illumina NextSeq. The figure shows binding across AXIN2, a known target gene of β-Catenin (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.

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IF-IC Image 7

Confocal immunofluorescent analysis of HeLa (left) and NCI-H28 (right) cells using β-Catenin (D10A8) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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Western Blotting Image 8

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

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Western Blotting Image 9

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

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Western Blotting Image 10

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

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Western Blotting Image 11

Western blot analysis of extracts from A204, SKMEL5, and NIH/3T3 cells using Slug (C19G7) Rabbit mAb.

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Western Blotting Image 12

Western blot analysis of extracts from COS cells, mock transfected or transfected with a construct expressing human TCF8/ZEB1, using TCF8/ZEB1 (D80D3) Rabbit mAb.

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Western Blotting Image 13

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

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Western Blotting Image 14

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

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IHC-P (paraffin) Image 15

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Vimentin (D21H3) XP® Rabbit mAb.

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IP Image 16

Immunoprecipitation of claudin-1 from A-431 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Claudin-1 (D5H1D) XP® Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot was performed using Claudin-1 (D5H1D) Rabbit mAb.

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Chromatin IP Image 17

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HCT 116 cells and either 20 μl of β-Catenin (D10A8) XP® Rabbit mAb or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human Axin2 Intron 1 Primers #8973, SimpleChIP® Human CaMK2D Intron 3 Primers #5111, human c-Myc promoter primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

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IHC-P (paraffin) Image 18

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using β-Catenin (D10A8) XP® Rabbit mAb.

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IP Image 19

Immunoprecipitation and western blot analysis of extracts from Hep G2 cells using ZO-1 (D7D12) Rabbit mAb. Lane 1 is 10% input.

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Flow Cytometry Image 20

Flow cytometric analysis of PANC-1 cells (blue) and A204 cells (green) using Slug (C19G7) Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

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Western Blotting Image 21

Western blot analysis of extracts from Jurkat, HT1080 and A172 cells using TCF8/ZEB1 (D80D3) Rabbit mAb.

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IHC-P (paraffin) Image 22

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using E-Cadherin (24E10) Rabbit mAb.

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IHC-P (paraffin) Image 23

Immunohistochemical analysis of paraffin-embedded human tonsil using Vimentin (D21H3) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).

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IHC-P (paraffin) Image 24

Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma using N-Cadherin (D4R1H) XP® Rabbit mAb.

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IHC-P (paraffin) Image 25

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Claudin-1 (D5H1D) XP® Rabbit mAb.

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IHC-P (paraffin) Image 26

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using β-Catenin (D10A8) XP® Rabbit mAb.

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IF-IC Image 27

Confocal immunofluorescent analysis of A204 cells (left) and PANC-1 cells (right) using Slug (C19G7) Rabbit mAb (green). Actin filaments have been labeled with DY554 phalloidin (red).

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IHC-P (paraffin) Image 28

Immunohistochemical analysis of paraffin-embedded human metastatic adenocarcinoma in lymph node, using E-Cadherin (24E10) Rabbit mAb.

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IHC-P (paraffin) Image 29

Immunohistochemical analysis of paraffin-embedded mouse colon using Vimentin (D21H3) XP® Rabbit mAb.

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IHC-P (paraffin) Image 30

Immunohistochemical analysis of paraffin-embedded A172 (positive, left) and MCF7 (negative, right) cell pellets using N-Cadherin (D4R1H) XP® Rabbit mAb.

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IHC-P (paraffin) Image 31

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Claudin-1 (D5H1D) XP® Rabbit mAb.

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IHC-P (paraffin) Image 32

Immunohistochemical analysis of paraffin-embedded cell pellets, HeLa (left) or NCI-H28 (right), using β-Catenin (D10A8) XP® Rabbit mAb.

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IHC-P (paraffin) Image 33

Immunohistochemical analysis of paraffin-embedded mouse lung using E-Cadherin (24E10) Rabbit mAb.

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Flow Cytometry Image 34

Flow cytometric analysis of HeLa cells, using Vimentin (D21H3) XP® Rabbit mAb (blue) compared to Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red).

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IF-IC Image 35

Confocal immunofluorescent analysis of A172 (positive, left) and MCF7 (negative, right) cells using N-Cadherin (D4R1H) XP® Rabbit mAb (green). Blue pseudocolor= DRAQ5® #4084 (fluorescent DNA dye).

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IHC-P (paraffin) Image 36

Immunohistochemical analysis of paraffin-embedded cell pellets, A-431 (left) and MCF7 (right), using Claudin-1 (D5H1D) XP® Rabbit mAb.

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IHC-P (paraffin) Image 37

Immunohistochemical analysis of paraffin-embedded mouse colon using β-Catenin (D10A8) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).

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IHC-P (paraffin) Image 38

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using E-Cadherin (24E10) Rabbit mAb in the presence of control peptide (left) or E-Cadherin Blocking Peptide #1050 (right).

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IHC-P (paraffin) Image 39

Immunohistochemical analysis of paraffin-embedded human skin using Claudin-1 (D5H1D) XP® Rabbit mAb.

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IHC-P (paraffin) Image 40

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using β-Catenin (D10A8) XP® Rabbit mAb.

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IHC-F (frozen) Image 41

Immunohistochemical analysis of frozen HCC827 xenograft, showing membrane and cytoplasmic localization using E-Cadherin (24E10) Rabbit mAb.

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IHC-F (frozen) Image 42

Immunohistochemical analysis of frozen mouse colon using β-Catenin (D10A8) XP® Rabbit mAb.

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Flow Cytometry Image 43

Flow cytometric analysis of HeLa cells (blue) and MCF7 cells (green) using E-Cadherin (24E10) Rabbit mAb.

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Flow Cytometry Image 44

Flow cytometric analysis of NCI-H28 (blue) or HeLa (green) cells using β-Catenin (D10A8) XP® Rabbit mAb.

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IF-IC Image 45

Confocal immunofluorescent images of MCF7 cells using E-Cadherin (24E10) Rabbit mAb (green, left) compared to an isotype control (right). Blue pseudocolor = DRAQ5® (fluorescent DNA dye).

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IF-F Image 46

Confocal immunofluorescent analysis of mouse colon using β-Catenin (D10A8) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Vimentin (D21H3) XP® Rabbit mAb 5741 20 µl
  • WB
  • IHC
  • IF
  • F
H M R Mk 57 Rabbit IgG
N-Cadherin (D4R1H) XP® Rabbit mAb 13116 20 µl
  • WB
  • IP
  • IHC
  • IF
H M 140 Rabbit IgG
Claudin-1 (D5H1D) XP® Rabbit mAb 13255 20 µl
  • WB
  • IP
  • IHC
H Dg 20 Rabbit IgG
β-Catenin (D10A8) XP® Rabbit mAb 8480 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
  • ChIP
H M R Mk 92 Rabbit IgG
ZO-1 (D7D12) Rabbit mAb 8193 20 µl
  • WB
  • IP
H Mk 220 Rabbit IgG
Snail (C15D3) Rabbit mAb 3879 20 µl
  • WB
  • IP
H M R Mk 29 Rabbit 
Slug (C19G7) Rabbit mAb 9585 20 µl
  • WB
  • IP
  • IF
  • F
H M 30 Rabbit IgG
TCF8/ZEB1 (D80D3) Rabbit mAb 3396 20 µl
  • WB
  • IP
H 200 Rabbit IgG
E-Cadherin (24E10) Rabbit mAb 3195 20 µl
  • WB
  • IHC
  • IF
  • F
H M 135 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

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

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 (D4R1H) XP® Rabbit mAb recognizes endogenous levels of total N-cadherin protein. Claudin-1 (D5H1D) XP® Rabbit mAb recognizes endogenous levels of total claudin-1 protein. 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. ß-Catenin (D10A8) XP® Rabbit mAb detects endogenous levels of total ß-catenin protein.

Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Arg526 of human N-cadherin protein, a synthetic peptide corresponding to residues near the carboxy terminus of human claudin-1 protein, 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.

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. Wheelock, M.J. and Johnson, K.R. (2003) Annu Rev Cell Dev Biol 19, 207-35.
  3. Cadigan, K.M. and Nusse, R. (1997) Genes Dev 11, 3286-305.
  4. Peinado, H. et al. (2007) Nat Rev Cancer 7, 415-28.
  5. Christofori, G. (2003) EMBO J 22, 2318-23.
  6. Wodarz, A. and Nusse, R. (1998) Annu Rev Cell Dev Biol 14, 59-88.
  7. Moreno-Bueno, G. et al. (2008) Oncogene 27, 6958-69.
  8. Hazan, R.B. et al. (2004) Ann N Y Acad Sci 1014, 155-63.
  9. Polakis, P. (1999) Curr Opin Genet Dev 9, 15-21.
  10. Helfand, B.T. et al. (2004) J Cell Sci 117, 133-41.
  11. Shin, K. et al. (2006) Annu Rev Cell Dev Biol 22, 207-35.
  12. Oliveira, S.S. and Morgado-Díaz, J.A. (2007) Cell Mol Life Sci 64, 17-28.
  13. Matter, K. and Balda, M.S. (2007) J Cell Sci 120, 1505-11.
  14. Hernandez, S. et al. (2007) Exp Cell Res 313, 1533-47.
  15. Umeda, K. et al. (2006) Cell 126, 741-54.
  16. Reichert, M. et al. (2000) J Biol Chem 275, 9492-500.
  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.
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.

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