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8655
Wnt/β-Catenin Activated Targets Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

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

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other Image 1 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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

other Image 2 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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

other Image 3 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

Chromatin immunoprecipitations were performed with cross-linked chromatin from 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 using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across Dclk1, a known target gene of c-Jun (see additional figure containing ChIP-qPCR data).

other Image 4 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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

other Image 5 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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).

other Image 6 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using Met (D1C2) XP® Rabbit mAb performed on the Leica® Bond™ Rx.

other Image 7 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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

other Image 8 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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

other Image 9 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

Chromatin immunoprecipitations were performed with cross-linked chromatin from mouse thymocytes and either TCF1/TCF7 (C63D9) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR using SimpleChIP® Mouse LEF1 Upstream Primers #80993 and SimpleChIP® Mouse MYT-1 Promoter Primers #8985. 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.

other Image 10 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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.

other Image 11 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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

other Image 12 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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

other Image 13 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

Chromatin immunoprecipitations were performed with cross-linked chromatin from 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 using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across chromosome 2 (upper), including Dclk1 (lower), a known target gene of c-Jun (see additional figure containing ChIP-qPCR data).

other Image 14 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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

other Image 15 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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

other Image 16 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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.

other Image 17 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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).

other Image 18 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

Flow cytometric analysis of A20 cells (blue) and EL-4 cells (green) using TCF1/TCF7 (C63D9) 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.

other Image 19 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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).

other Image 20 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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).

other Image 21 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

Chromatin immunoprecipitations were performed with cross-linked chromatin from PC-12 cells starved overnight and treated with β-NGF #5221 (50ng/ml) for 2h and either of c-Jun (60A8) 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® 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.

other Image 22 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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

other Image 23 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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

other Image 24 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

Western blot analysis of extracts from control HEK293 cells (lane 1) or c-Myc knockout HEK293 cells (lane 2) using c-Myc (D84C12) Rabbit mAb Antibody, #5605 (upper) or β-actin (13E5) Rabbit mAb, #4970 (lower). The absence of signal in the c-Myc knockout HEK293 cells confirms specificity of the antibody for c-Myc.

other Image 25 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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).

other Image 26 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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

other Image 27 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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

other Image 28 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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.

other Image 29 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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

other Image 30 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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.

other Image 31 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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

other Image 32 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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

other Image 33 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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

other Image 34 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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

other Image 35 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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

other Image 36 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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

other Image 37 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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).

other Image 38 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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

other Image 39 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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

other Image 40 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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

other Image 41 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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

other Image 42 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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

other Image 43 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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

other Image 44 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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

other Image 45 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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

other Image 46 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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).

other Image 47 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

Western blot analysis of extracts from control HeLa cells (lane 1) or c-Jun knockout HeLa cells (lane 2) using c-Jun (60A8) Rabbit mAb #9165. The absence of signal in the c-Jun knockout HeLa cells confirms specificity of the antibody for c-Jun.

other Image 48 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

Western blot analysis of extracts from control HeLa cells (lane 1) or Met knockout HeLa cells (lane 2) using Met (D1C2) XP® Rabbit mAb #8198. The absence of signal in the Met knockout HeLa cells confirms specificity of the antibody for Met.

other Image 49 - Wnt/β-Catenin Activated Targets Antibody Sampler Kit

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

To Purchase # 8655T
Product # Size Price
8655T
1 Kit  (8 x 20 µl) $ 554

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
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
  • ChIP
H M 48, 50 Rabbit IgG
Anti-mouse IgG, HRP-linked Antibody 7076 100 µl
  • WB
Horse 
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

Product Description

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.

Specificity / Sensitivity

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.

Source / Purification

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.

Background

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.

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.