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8655
Wnt/β-Catenin Activated Targets Antibody Sampler Kit

Wnt/β-Catenin Activated Targets Antibody Sampler Kit #8655

Western Blotting Image 1

Western blot analysis of extracts from HeLa and PANC1 cell lines using CD44 (156-3C11) Mouse mAb.

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Western Blotting Image 2

Western blot analysis of extracts from MCF7, L929 and C6 cells, using Cyclin D1 (92G2) Rabbit mAb.

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Chromatin IP-seq Image 3

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 PC-12 cells starved overnight and treated with β-NGF #5221 (50ng/ml) for 2h and c-Jun (60A8) Rabbit mAb, using SimpleChIP® Plus 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 Dclk1, a known target gene of c-Jun (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.

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Western Blotting Image 4

Western blot analysis of extracts from NIH/3T3 and SK-N-MC cells, untreated or UV-treated, using c-Jun (60A8) Rabbit mAb.

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Western Blotting Image 5

Western blot analysis of total cell lysates from HCT15, DLD1 and mouse thymocytes using LEF1 (C12A5) Rabbit mAb.

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Western Blotting Image 6

Western blot analysis of extracts from HT-29 (Met+), SK-BR-3 (Met-), and T-47D (Met-) cells using Met (D1C2) XP® Rabbit mAb (upper) or β-Actin Antibody #4967 (lower).

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Western Blotting Image 7

Western blot analysis of extracts mouse intestine (MMP-7 positive), mouse colon (MMP-7 negative) and rat prostate (MMP-7 positive) tissues using MMP-7 (D4H5) XP® Rabbit mAb.

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Western Blotting Image 8

Western blot analysis of extracts from HeLa cells, mock transfected or transfected with SignalSilence® c-Myc siRNA I #6341, using c-Myc (D84C12) Rabbit mAb.

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Western Blotting Image 9

Western blot analysis of total cell lysates from HT29, Colo201, Jurkat and mouse thymocytes using TCF1/TCF7 (C63D9) Rabbit mAb.

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Western Blotting Image 10

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 11

Immunohistochemical analysis of human Non-Hodgkin's lymphoma using CD44 (156-3C11) Mouse mAb.

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IHC-P (paraffin) Image 12

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using Cyclin D1 (92G2) Rabbit mAb.

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IHC-P (paraffin) Image 13

Immunohistochemical analysis of paraffin-embedded human astrocytoma, using c-Jun (60A8) Rabbit mAb.

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Chromatin IP Image 14

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 PC-12 cells starved overnight and treated with β-NGF #5221 (50ng/ml) for 2h and either 10 μl of c-Jun (60A8) 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® Rat CCRN4L Promoter Primers #7983, rat DCLK1 promoter primers, and SimpleChIP® Rat GAPDH Promoter Primers #7964. 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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Flow Cytometry Image 15

Flow cytometric analysis of untreated Jurkat cells using LEF1 (C12A5) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).

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IHC-P (paraffin) Image 16

Immunohistochemical analysis of paraffin-embedded human metastatic lung carcinoma using Met (D1C2) XP® Rabbit mAb.

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Western Blotting Image 17

Western blot analysis of extracts from COS cells, untransfected or transfected with mouse MMP-7, using MMP-7 (D4H5) XP® Rabbit mAb.

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Western Blotting Image 18

Western blot analysis of extracts from various cell lines using c-Myc (D84C12) Rabbit mAb.

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IHC-P (paraffin) Image 19

Immunohistochemical analysis of paraffin-embedded human Non-Hodgkin's lymphoma using TCF1/TCF7 (C63D9) Rabbit mAb.

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IHC-P (paraffin) Image 20

Immunohistochemical analysis of human tonsil using CD44 (156-3C11) Mouse mAb.

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IHC-P (paraffin) Image 21

Immunohistochemical analysis of paraffin-embedded Apc (min/+) mouse intestine using Cyclin D1 (92G2) Rabbit mAb.

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IHC-P (paraffin) Image 22

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using c-Jun (60A8) Rabbit mAb.

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IF-IC Image 23

Confocal immunofluorescent analysis of HCT-15 cells using LEF1 (C12A5) Rabbit mAb (green). Actin filaments have been labeled with DyLight™ 554 Phalloidin #13054 (red).

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IHC-P (paraffin) Image 24

Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma using Met (D1C2) XP® Rabbit mAb.

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IHC-P (paraffin) Image 25

Immunohistochemical analysis of paraffin-embedded mouse small intestine (positive, left) and mouse colon (negative, right) using MMP-7 (D4H5) XP® Rabbit mAb.

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Flow Cytometry Image 26

Flow cytometric analysis of Raji cells using c-Myc (D84C12) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).

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IHC-P (paraffin) Image 27

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using TCF1/TCF7 (C63D9) Rabbit mAb in the presence of control peptide (left) or TCF1/TCF7 blocking peptide #1007 (right).

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Flow Cytometry Image 28

Flow cytometric analysis of Hela cells using CD44 (156-3C11) Mouse mAb (blue) compared to a nonspecific negative control antibody (red).

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IHC-P (paraffin) Image 29

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Cyclin D1 (92G2) Rabbit mAb in the presence of control peptide (left) or Cyclin D1 Blocking Peptide #1044 (right).

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IHC-P (paraffin) Image 30

Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma, using c-Jun (60A8) Rabbit mAb.

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IHC-P (paraffin) Image 31

Immunohistochemical analysis of paraffin-embedded human papillary renal cell carcinoma using Met (D1C2) XP® Rabbit mAb.

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IF-IC Image 32

Confocal immunofluorescent analysis of HeLa cells, mock-transfected (left) or transfected with SignalSilence® c-Myc siRNA I #6341 (right), using c-Myc (D84C12) Rabbit mAb (green). Actin filaments have been labeled wth DY-554 phalloidin (red).

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IHC-P (paraffin) Image 33

Immunohistochemical analysis of paraffin-embedded human tonsil using TCF1/TCF7 (C63D9) Rabbit mAb.

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Flow Cytometry Image 34

Flow cytometric analysis of whole blood using CD44 (156-3C11) Mouse mAb.

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IHC-P (paraffin) Image 35

Immunohistochemical analysis of paraffin-embedded H1975 xenograft, using Cyclin D1 (92G2) Rabbit mAb.

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Flow Cytometry Image 36

Flow cytometric analysis of Jurkat cells using c-Jun (60A8) Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

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IHC-P (paraffin) Image 37

Immunohistochemical analysis of paraffin-embedded cell pellets, MKN-45 (left) and T-47D (right), using Met (D1C2) XP® Rabbit mAb.

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Flow Cytometry Image 38

Flow cytometric analysis of Jurkat cells using TCF1/TCF7 (C63D9) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).

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IF-IC Image 39

Confocal immunofluorescent analysis of Hela cells using CD44 (156-3C11) Mouse mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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Flow Cytometry Image 40

Flow cytometric analysis of untreated HT29 cells, using Cyclin D1 (92G2) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).

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IF-IC Image 41

Confocal immunofluorescent analysis of HeLa cells, using c-Jun (60A8) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).

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IHC-F (frozen) Image 42

Immunohistochemical analysis of frozen MKN-45 xenograft using Met (D1C2) XP® Rabbit mAb.

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IF-IC Image 43

Confocal immunofluorescent analysis of DLD-1 cells using TCF1/TCF7 (C63D9) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).

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Flow Cytometry Image 44

Flow cytometric analysis of T-47D cells (blue) and HT-29 cells (green) using Met (D1C2) XP® Rabbit mAb.

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IF-IC Image 45

Confocal immunofluorescent analysis of HT-29 and T-47D cells using Met (D1C2) 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
CD44 (156-3C11) Mouse mAb 3570 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H 80 Mouse IgG2a
Cyclin D1 (92G2) Rabbit mAb 2978 20 µl
  • WB
  • IHC
  • F
H M R 36 Rabbit IgG
c-Jun (60A8) Rabbit mAb 9165 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
  • ChIP
H M R Mk 43, 48 Rabbit IgG
LEF1 (C12A5) Rabbit mAb 2230 20 µl
  • WB
  • IF
  • F
H M R 25-58 Rabbit IgG
Met (D1C2) XP® Rabbit mAb 8198 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H 140, 170 Rabbit IgG
MMP-7 (D4H5) XP® Rabbit mAb 3801 20 µl
  • WB
  • IHC
M R 28, 20-22 Rabbit 
c-Myc (D84C12) Rabbit mAb 5605 20 µl
  • WB
  • IF
  • F
H M R 57-65 Rabbit IgG
TCF1/TCF7 (C63D9) Rabbit mAb 2203 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H M 48, 50 Rabbit 
Anti-mouse IgG, HRP-linked Antibody 7076 100 µl
  • WB
Horse 
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

The Wnt/β-Catenin Activated Targets Antibody Sampler Kit provides an economical means to investigate target proteins of the Wnt/β-Catenin signaling pathway. This kit contains enough primary antibody to perform two western blots per primary.

Each antibody in the Wnt/β-Catenin Activated Targets Antibody Sampler Kit detects its respective target at endogenous levels. LEF1 (C12A5) Rabbit mAb does not recognize the dominant negative forms of LEF1 generated by an alternative promoter. c-Myc (D84C12) Rabbit mAb is not recommended for detection of Myc-tagged fusion proteins. TCF1/TCF7 (C63D9) Rabbit mAb does not recognize the dominant negative isoforms of TCF1/TCF7 lacking the amino-terminal β-catenin binding domain and does not cross-react with LEF1.

Rabbit monoclonal antibodies are produced by immunizing animals with a GST-c-Jun protein corresponding to the amino-terminal sequence of human c-Jun protein or synthetic peptides corresponding to residues near the amino terminus of c-Myc protein, residues surrounding Pro82 of human LEF1 protein, residues surrounding Ile264 of mouse MMP-7 protein, residues surrounding Pro95 of human TCF1/TCF7 protein, residues near the carboxy terminus of human Met protein, or residues near the carboxy-terminus of human cyclin D1 protein.

The mouse monoclonal antibody is produced by immunizing BALB/c mice with stimulated human leukocytes for CD44.

The Wnt family includes several secreted glycoproteins that play important roles in animal development. β-catenin is a key downstream effector of the Wnt signaling pathway that research studies have shown is implicated in early embryonic development and tumorigenesis in vertebrates (1-3). Following binding of Wnt family proteins to the Frizzled receptor, β-catenin translocates to the nucleus where it interacts with LEF1 and TCF1 to activate canonical targets (4). Accepted canonical targets include CD44, cyclin D1, c-Jun, c-Myc, Met, and MMP-7 (5-11).

  1. Cadigan, K.M. and Nusse, R. (1997) Genes Dev 11, 3286-305.
  2. Wodarz, A. and Nusse, R. (1998) Annu Rev Cell Dev Biol 14, 59-88.
  3. Polakis, P. (1999) Curr Opin Genet Dev 9, 15-21.
  4. Brantjes, H. et al. (2002) Biol Chem 383, 255-61.
  5. Zeilstra, J. et al. (2008) Cancer Res 68, 3655-61.
  6. Shtutman, M. et al. (1999) Proc Natl Acad Sci U S A 96, 5522-7.
  7. Mann, B. et al. (1999) Proc Natl Acad Sci U S A 96, 1603-8.
  8. Wilkins, J.A. and Sansom, O.J. (2008) Cancer Res 68, 4963-6.
  9. Boon, E.M. et al. (2002) Cancer Res 62, 5126-8.
  10. Brabletz, T. et al. (1999) Am J Pathol 155, 1033-8.
  11. Clevers, H. and Nusse, R. (2012) Cell 149, 1192-205.
For Research Use Only. Not For Use In Diagnostic Procedures.

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