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9383
TCF/LEF Family Antibody Sampler Kit

TCF/LEF Family Antibody Sampler Kit #9383

Western Blotting Image 1

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 2

Western blot analysis of total lysates from various cell lines using TCF3/TCF7L1 (D15G11) Rabbit mAb.

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Western Blotting Image 3

Western blot analysis of extracts from various cell types using TCF4/TCF7L2 (C48H11) Rabbit mAb.

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Chromatin IP-seq Image 4

Chromatin immunoprecipitations were performed with cross-linked chromatin from HCT116 cells and either TCF4/TCF7L2 (C48H11) Rabbit mAb or Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb #8814, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. TCF4/TCF7L2 and β-Catenin are known to associate with each other on chromatin. The figure shows binding of both TCF4/TCF7L2 and β-Catenin across CAMK2D, a known target gene of both TCF4/TCF7L2 and β-Catenin (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 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

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 7

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

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Chromatin IP Image 8

Chromatin immunoprecipitations were performed with cross-linked chromatin from HCT 116 cells and either TCF4/TCF7L2 (C48H11) 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 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.

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Flow Cytometry Image 9

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 10

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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IF-IC Image 11

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 12

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

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Flow Cytometry Image 13

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 14

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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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
TCF1/TCF7 (C63D9) Rabbit mAb 2203 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H M 48, 50 Rabbit 
TCF3/TCF7L1 (D15G11) Rabbit mAb 2883 20 µl
  • WB
H Mk 78 Rabbit 
TCF4/TCF7L2 (C48H11) Rabbit mAb 2569 20 µl
  • WB
  • IP
  • ChIP
H 58, 79 Rabbit IgG
LEF1 (C12A5) Rabbit mAb 2230 20 µl
  • WB
  • IF
  • F
H M R 25-58 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

The TCF/LEF Family Antibody Sampler Kit provides an economical means of detecting total protein from the TCF/LEF1 family members. The kit contains enough primary and secondary antibodies to perform two Western blots with each antibody.

Each antibody in this kit recognizes endogenous levels of total protein for the specified target and does not cross-react with other family members. TCF1/TCF7 (C63D9) Rabbit mAb does not recognize the dominant negative isoforms of TCF1/TCF7 lacking the amino-terminal β-catenin binding domain. LEF1 (C12A5) Rabbit mAb does not recognize the dominant negative forms of LEF1 generated by an alternative promoter.

Monoclonal antibody is produced by immunizing animals with synthetic peptides corresponding to regions surrounding Pro95 of human TCF1/TCF7 protein, Gln96 of human TCF3/TCF7L1, Leu330 of human TCF4/TCF7L2, and Pro82 of human LEF1.

LEF1 and TCF are members of the high mobility group (HMG) DNA binding protein family of transcription factors which consists of the following: Lymphoid Enhancer Factor 1 (LEF1), T Cell Factor 1 (TCF1, also known as TCF7), TCF3 (also known as TCF7L1) and TCF4 (also known as TCF7L2) (1). LEF1 and TCF1/TCF7 were originally identified as important factors regulating early lymphoid development (2) that act downstream in Wnt signaling. LEF1 and TCF bind to Wnt response elements to provide docking sites for β-catenin, which translocates to the nucleus to promote the transcription of target genes upon activation of Wnt signaling (3). LEF1 and TCF proteins are dynamically expressed during development and aberrant activation of the Wnt signaling pathway is involved in many types of cancers including colon cancer (4,5).

  1. Waterman, M.L. (2004) Cancer Metastasis Rev 23, 41-52.
  2. Schilham, M.W. and Clevers, H. (1998) Semin Immunol 10, 127-32.
  3. Brantjes, H. et al. (2002) Biol Chem 383, 255-61.
  4. Reya, T. and Clevers, H. (2005) Nature 434, 843-50.
  5. Logan, C.Y. and Nusse, R. (2004) Annu Rev Cell Dev Biol 20, 781-810.
Entrez-Gene Id
51176 , 83439 , 6934 , 6932
Swiss-Prot Acc.
Q9UJU2 , Q9HCS4 , Q9NQB0 , P36402
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.

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