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9383
TCF/LEF Family Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit
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TCF/LEF Family Antibody Sampler Kit #9383

Citations (10)
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 (0.1 mg/mL) from HepG2 cells using TCF4/TCF7L2 (C48H11) Rabbit mAb #2569. The virtual lane view (left) shows the target bands (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 various cell types using TCF4/TCF7L2 (C48H11) Rabbit mAb.
CUT&Tag was performed with HCT 116 cells and TCF4/TCF7L2 (C48H11) Rabbit mAb, using CUT&Tag Assay Kit #77552. DNA library was prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figure shows binding across MYC, a known target gene of TCF4 (see our ChIP-qPCR figure).
Western blot analysis of total lysates from various cell lines using TCF3/TCF7L1 (D15G11) Rabbit mAb.
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.
Immunohistochemical analysis of paraffin-embedded human Non-Hodgkin lymphoma using TCF1/TCF7 (C63D9) Rabbit mAb.
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 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #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 datasheet.
CUT&Tag was performed with HCT116 cells and TCF4/TCF7L2 (C48H11) Rabbit mAb, using CUT&Tag Assay Kit #77552. DNA library was prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figures show binding across chromosome 8 (upper), including MYC (lower), a known target gene of TCF4 (see our ChIP-qPCR figure).
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).
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 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #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 chromosome 4 (upper), including CAMK2D (lower), a known target gene of both TCF4/TCF7L2 and β-Catenin (see additional figure containing ChIP-qPCR data).
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).
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.
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.

CUT&RUN was performed with HCT 116 cells and TCF4/TCF7L2 (C48H11) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figures show binding across MYC, a known target gene of TCF4 (see additional figure containing CUT&RUN-qPCR data).
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.
CUT&RUN was performed with HCT 116 cells and TCF4/TCF7L2 (C48H11) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 8 (upper), including MYC (lower), a known target gene of TCF4 (see additional figure containing CUT&RUN-qPCR data).
CUT&RUN was performed with HCT 116 cells and either TCF4/TCF7L2 (C48H11) Rabbit mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using human PRKACB intron 1 primers, human MYC exon 1 primers, and human ITM2A upstream primers. 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.
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.
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To Purchase # 9383
Cat. # Size Qty. Price
9383T		 1 Kit  (4 x 20 microliters)

Bulk & Custom Formulation

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
TCF1/TCF7 (C63D9) Rabbit mAb 2203 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
  • ChIP
 H M 48, 50 Rabbit IgG
TCF3/TCF7L1 (D15G11) Rabbit mAb 2883 20 µl
  • WB
 H Mk 78 Rabbit 
TCF4/TCF7L2 (C48H11) Rabbit mAb 2569 20 µl
  • WB
  • IP
  • ChIP
  • C&R
  • C&T
 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 

Product Description

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.

Specificity / Sensitivity

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.

Source / Purification

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.

Background

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).

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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