Cell Signaling Technology

Product Pathways - Development

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

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
CD44 (156-3C11) Mouse mAb #3570 40 µl W IP IHC-P IF-IC F H 80 Mouse IgG2a
Cyclin D1 (DCS6) Mouse mAb #2926 40 µl W IP H M R 36 Mouse IgG2a
c-Jun (60A8) Rabbit mAb #9165 40 µl W IP IHC-P IHC-F IF-IC ChIP H M R Mk 43, 48 Rabbit IgG
LEF1 (C12A5) Rabbit mAb #2230 40 µl W IP IF-IC F H M R 25-58 kDa Rabbit IgG
Met (D1C2) XP® Rabbit mAb #8198 40 µl W IP IHC-P IHC-F IF-IC F H 140, 170 Rabbit IgG
MMP-7 (D4H5) XP® Rabbit mAb #3801 40 µl W IHC-P M R 28, 20-22 Rabbit
c-Myc (D84C12) XP® Rabbit mAb #5605 40 µl W IP IF-IC F H M R (Mk) (Dg) (Pg) 57-65 Rabbit IgG
TCF1 (C63D9) Rabbit mAb #2203 40 µl W IP IHC-P IF-IC F H M 48, 50 kDa Rabbit IgG
Anti-mouse IgG, HRP-linked Antibody #7076 100 µl Horse
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-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry  ChIP=Chromatin IP
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  Dg=Dog  Pg=Pig
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

Cyclin D1 (DCS6) Mouse mAb recognizes endogenous levels of total cyclin D1 protein. This antibody does not cross-react with cyclin D2 or D3. c-Jun (60A8) Rabbit mAb recognizes endogenous levels of total c-Jun protein, regardless of phosphorylation state. LEF1 (C12A5) Rabbit mAb recognizes endogenous levels of total LEF1 protein. This antibody does not recognize the dominant negative forms of LEF1 generated by an alternative promoter. c-Myc (D84C12) XP® Rabbit mAb recognizes endogenous levels of total c-Myc protein. This antibody is not recommended for detection of Myc-tagged fusion proteins. TCF1 (C63D9) Rabbit mAb recognizes endogenous levels of total TCF1 protein. This antibody does not recognize the dominant negative isoforms of TCF1 lacking the amino-terminal β-catenin binding domain and does not cross-react with LEF1. CD44 (156-3C11) Mouse mAb, Met (D1C2) XP® Rabbit mAb, and MMP-7 (D4H5) XP® Rabbit mAb all recognize endogenous levels of total respective protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from HCT29 cells, Colo201 cells, Jurkat cells, and mouse thymocytes using TCF1 (C63D9) Rabbit mAb #2203.

Western Blotting

Western Blotting

Western blot analysis of extracts from HCT15 cells, DLD1 cells, and mouse thymocytes using LEF1 (C12A5) Rabbit mAb #2230.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa and 3T3 cells using Cyclin D1 (DCS6) Mouse mAb #2926.


Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa and PANC1 cells using CD44 (156-3C11) Mouse mAb #3570.

Western Blotting

Western Blotting

Western blot analysis of extracts from mouse intestine (MMP-7-positive), mouse colon (MMP-7-negative), and rat prostate (MMP-7-positive) tissues using MMP-7 (D4H5) XP® Rabbit mAb #3801.

Western Blotting

Western Blotting

Western blot analysis of extracts from Jurkat, RPMI 8226, and A20 cells using c-Myc (D84C12) XP® Rabbit mAb #5605.


Western Blotting

Western Blotting

Western blot analysis of extracts from HT-29 (Met-positive), SK-BR-3 (Met-negative), and T-47D (Met-negative) cells using Met (D1C2) XP® Rabbit mAb #8198 (upper) or β-Actin Antibody #4967 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from NIH/3T3 and SK-N-MC cells, untreated (-) or UV-treated (+), using c-Jun (60A8) Rabbit mAb #9165.

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 four western blots per primary.

Source / Purification

Mouse monoclonal antibodies are produced by immunizing BALB/c mice with stimulated human leukocytes for CD44 or immunizing animals with a recombinant human cyclin D1 corresponding to residues 151-170.Rabbit monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro82 of human LEF1 protein, residues near the carboxy terminus of human Met protein, residues surrounding Ile264 of mouse MMP-7 protein, amino-terminal residues of c-Myc protein, a region surrounding Pro95 of human TCF1 protein, or a GST-c-Jun protein corresponding to the amino-terminal sequence of human c-Jun protein.

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.

Application References

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

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