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9782
Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

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

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Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 1

Flow cytometric analysis of MCF7 cells (blue) and HeLa cells (green) using Vimentin (D21H3) XP® 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.

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 2

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

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 3

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

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 4

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).

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 5

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

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 6

Chromatin immunoprecipitations were performed with cross-linked chromatin from HCT116 cells and either β-Catenin (D10A8) XP® Rabbit mAb or 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 using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. 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.

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: 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).

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 8

Western blot analysis of extracts from control HeLa cells (lane 1) or HeLa cells with an apparent in-frame truncation mutation in the gene encoding β-Catenin (lane 2) using β-Catenin (D10A8) XP® Rabbit mAb, #8480 (upper) or β-actin (D6A8) Rabbit mAb #8457 (lower). The change in β-Catenin molecular weight in the mutated HeLa cells is consistent with an in-frame deletion.

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 9

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

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 10

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

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 11

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.

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 12

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

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 13

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

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: 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.

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 15

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

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 16

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

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 17

Immunohistochemical analysis of paraffin-embedded human non-Hodgkin’s Lymphoma using N-Cadherin (D4R1H) XP® Rabbit mAb performed on the Leica® BOND™ Rx.

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 18

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

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 19

Chromatin immunoprecipitations were performed with cross-linked chromatin from HCT 116 cells and either β-Catenin (D10A8) XP® Rabbit mAb or 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.

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 20

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

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 21

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).

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 22

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

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 23

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).

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 24

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

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 25

Immunohistochemical analysis of paraffin-embedded human granulosa cell tumor of the ovary using N-Cadherin (D4R1H) XP® Rabbit mAb performed on the Leica® BOND™ Rx.

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 26

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

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 27

Flow cytometric analysis of NCI-H28 cells (blue) and HeLa cells (green) using β-Catenin (D10A8) XP® Rabbit mAb (solid lines) or concentration-matched Rabbit Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as secondary antibody.

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 28

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

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 29

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

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 30

Immunohistochemical analysis of paraffin-embedded human gastric carcinoma using N-Cadherin (D4R1H) XP® Rabbit mAb performed on the Leica® BOND™ Rx.

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 31

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

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 32

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).

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 33

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

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 34

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).

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 35

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.

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 36

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

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 37

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

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 38

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).

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 39

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

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 40

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

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 41

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

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 42

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

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 43

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).

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 44

Western blot analysis of extracts from control HeLa cells (lane 1) or Vimentin knockout HeLa cells (lane 2) using Vimentin (D21H3) XP® Rabbit mAb #5741 (upper) or β-Actin (13E5) Rabbit mAb #4970 (lower). The absence of signal in the Vimentin knockout HeLa cells confirms specificity of the antibody for Vimentin.

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 45

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).

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 46

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

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 47

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

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 48

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

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 49

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).

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 50

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

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 51

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

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 52

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

Epithelial-Mesenchymal Transition (EMT) Antibody Sampler Kit: Image 53

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

To Purchase # 9782T
Product # Size Price
9782T
1 Kit  (9 x 20 µl) $ 609

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
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 

Product Description

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.

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 (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.

Source / Purification

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.

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, 3286-3305.
  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.

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