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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

Citations (9)
Confocal immunofluorescent analysis of fixed frozen lymph node from wild-type (left) Tcf7GFP flox (right; Jax Strain 030909) mice using TCF1/TCF7 (C63D9) Rabbit mAb #2203 (red) and CD4 (RM4-5) Rat mAb (redFluor 710 Conjugate) #75508 (blue). EGFP insertion around Tcf7 exon 2 interferes with expression of long isoforms, but not short isoforms.

Flow cytometric analysis of human peripheral blood mononuclear cells using LEF1 (C12A5) Rabbit mAb (right) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (left), co-stained with CD3 (UCHT1) Mouse mAb (APC Conjugate) #19881. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

Simple Western™ analysis of lysates (1 mg/mL) from Raji cells using c-Myc (D84C12) Rabbit mAb #5605. The virtual lane view (left) shows a single target band (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Simple Western™ analysis of lysates (0.1 mg/mL) from HT-29 untreated cells using Met (D1C2) XP® Rabbit mAb #8198 lane view (left) shows a single target band (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.

Simple Western™ analysis of lysates (0.1 mg/mL) from HeLa cells treated with UV (100 mJ/cm2; 2H recovery) using c-Jun (60A8) Rabbit mAb #9165. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ ​​​​​​​ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.

Western blot analysis of total cell lysates from HT29, Colo201, Jurkat and mouse thymocytes using TCF1/TCF7 (C63D9) Rabbit mAb.
Western blot analysis of total cell lysates from HCT15, DLD1 and mouse thymocytes using LEF1 (C12A5) Rabbit mAb.
Western blot analysis of extracts from MCF7, L929 and C6 cells, using Cyclin D1 (92G2) Rabbit mAb.
Western blot analysis of extracts from HeLa and PANC1 cell lines using CD44 (156-3C11) Mouse mAb.
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.
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.
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.
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.
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).
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.
Immunohistochemical analysis of paraffin-embedded human Non-Hodgkin lymphoma using TCF1/TCF7 (C63D9) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using Cyclin D1 (92G2) Rabbit mAb.
Immunohistochemical analysis of human Non-Hodgkin's lymphoma using CD44 (156-3C11) Mouse mAb.
Western blot analysis of extracts from COS cells, untransfected or transfected with mouse MMP-7, using MMP-7 (D4H5) XP® Rabbit mAb.
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.
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.
Western blot analysis of extracts from NIH/3T3 and SK-N-MC cells, untreated or UV-treated, using c-Jun (60A8) Rabbit mAb.
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).
Immunohistochemical analysis of paraffin-embedded Apc (min/+) mouse intestine using Cyclin D1 (92G2) Rabbit mAb.
Immunohistochemical analysis of human tonsil using CD44 (156-3C11) Mouse mAb.
Immunohistochemical analysis of paraffin-embedded mouse small intestine (positive, left) and mouse colon (negative, right) using MMP-7 (D4H5) XP® Rabbit mAb.
Western blot analysis of extracts from various cell lines using c-Myc (D84C12) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using Met (D1C2) XP® Rabbit mAb performed on the Leica® Bond Rx.
Immunohistochemical analysis of paraffin-embedded human astrocytoma, using c-Jun (60A8) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human tonsil using TCF1/TCF7 (C63D9) Rabbit mAb.
Confocal immunofluorescent analysis of HCT-15 cells using LEF1 (C12A5) Rabbit mAb (green). Actin filaments have been labeled with DyLight 554 Phalloidin #13054 (red).
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).
Confocal immunofluorescent analysis of PANC-1 cells (left, positive) or A2780 cells (right, negative) using CD44 (156-3C11) Mouse mAb (green), DyLight 650 Phalloidin #12956 (red), and DAPI #4083 (blue).
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).
Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using Met (D1C2) XP® Rabbit mAb performed on the Leica® Bond Rx.
Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using c-Jun (60A8) Rabbit mAb.
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).

Flow cytometric analysis of mouse splenocytes using LEF1 (C12A5) Rabbit mAb (right) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (left), co-stained with CD3 (17A2) Rat mAb (APC Conjugate) #24265. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

Immunohistochemical analysis of paraffin-embedded H1975 xenograft, using Cyclin D1 (92G2) Rabbit mAb.
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).
Immunohistochemical analysis of paraffin-embedded human metastatic lung carcinoma using Met (D1C2) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma, using c-Jun (60A8) Rabbit mAb.
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.
Flow cytometric analysis of live LNCaP cells (blue, negative) and PANC-1 cells (green, positive) using CD44 (156-3C11) Mouse mAb (solid lines) or a concentration-matched Mouse (G3A1) mAb IgG1 Isotype Control #5415 (dashed lines). Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 was used as a secondary antibody.
Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma using Met (D1C2) XP® Rabbit mAb.
Flow cytometric analysis of fixed and permeabilized LNCaP cells (blue, negative) and PANC-1 cells (green, positive) using CD44 (156-3C11) Mouse mAb (solid lines) or concentration-matched Mouse (G3A1) mAb IgG1 Isotype Control #5415 (dashed lines). Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 was used as a secondary antibody.
Immunohistochemical analysis of paraffin-embedded human papillary renal cell carcinoma using Met (D1C2) XP® Rabbit mAb.
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.
Immunohistochemical analysis of paraffin-embedded cell pellets, MKN-45 (left) and T-47D (right), using Met (D1C2) XP® Rabbit mAb.
Confocal immunofluorescent analysis of HeLa cells, using c-Jun (60A8) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).
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.
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).
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 Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across Dclk1, a known target gene of c-Jun (see additional figure containing ChIP-qPCR data).
Flow cytometric analysis of fixed and permeabilized Ramos cells (blue, negative) and MKN-45 cells (green, positive) using Met (D1C2) 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.
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 Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #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).
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.
To Purchase # 8655
Cat. # Size Qty. Price
8655T
1 Kit  (8 x 20 microliters)

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
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).

Limited Uses

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For Research Use Only. Not for Use in Diagnostic Procedures.
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U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
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