Recombinant
RabMAb

Recombinant Anti-LEF1 antibody [EPR2029Y] (ab137872)

Rabbit recombinant monoclonal LEF1 antibody [EPR2029Y]. Validated in WB, IP, IHC, Flow Cyt, ICC/IF and tested in Mouse, Rat, Human. Cited in 15 publication(s). Independently reviewed in 1 review(s).

Overview

  • Product name
    Anti-LEF1 antibody [EPR2029Y]
    See all LEF1 primary antibodies
  • Description
    Rabbit monoclonal [EPR2029Y] to LEF1
  • Host species
    Rabbit
  • Tested applications
    Suitable for: Flow Cyt, IP, WB, IHC-P, ICC/IF, IHC-FoFrmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human LEF1 aa 100-200. The exact sequence is proprietary.
    Database link: Q9UJU2

  • Positive control
    • WB: Jurkat whole cell lysate (ab7899); Rat thymus tissue lysate; Human fetal lysate; His-tagged mouse LEF-1 recombinant protein (aa1-397). IHC-P: Human tonsil and thymus tissues; Mouse and rat spleen tissues. ICC/IF: Jurkat cells. Flow Cyt: Jurkat cells.
  • General notes

      

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab137872 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
WB 1/1000. Predicted molecular weight: 44 kDa.

We don’t recommend this antibody for mouse in Western Blot. In our hands an extra band was observed in mouse tissue lysates.

IHC-P 1/100 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

ICC/IF 1/500.
IHC-FoFr Use at an assay dependent concentration. PubMed: 24586192

Target

  • Function
    Participates in the Wnt signaling pathway. Activates transcription of target genes in the presence of CTNNB1 and EP300. May play a role in hair cell differentiation and follicle morphogenesis. TLE1, TLE2, TLE3 and TLE4 repress transactivation mediated by LEF1 and CTNNB1. Regulates T-cell receptor alpha enhancer function. Binds DNA in a sequence-specific manner. PIAG antagonizes both Wnt-dependent and Wnt-independent activation by LEF1 (By similarity). Isoform 3 lacks the CTNNB1 interaction domain and may be an antagonist for Wnt signaling. Isoform 5 transcriptionally activates the fibronectin promoter, binds to and represses transcription from the E-cadherin promoter in a CTNNB1-independent manner, and is involved in reducing cellular aggregation and increasing cell migration of pancreatic cancer cells. Isoform 1 transcriptionally activates MYC and CCND1 expression and enhances proliferation of pancreatic tumor cells.
  • Tissue specificity
    Detected in thymus. Not detected in normal colon, but highly expressed in colon cancer biopsies and colon cancer cell lines. Expressed in several pancreatic tumors and weakly expressed in normal pancreatic tissue. Isoforms 1 and 5 are detected in several pancreatic cell lines.
  • Sequence similarities
    Belongs to the TCF/LEF family.
    Contains 1 HMG box DNA-binding domain.
  • Domain
    Proline-rich and acidic regions are implicated in the activation functions of RNA polymerase II transcription factors.
  • Cellular localization
    Nucleus. Found in nuclear bodies upon PIASG binding.
  • Information by UniProt
  • Database links
  • Alternative names
    • DKFZp586H0919 antibody
    • FLJ46390 antibody
    • LEF 1 antibody
    • LEF-1 antibody
    • Lef1 antibody
    • LEF1_HUMAN antibody
    • Lymphoid enhancer binding factor 1 antibody
    • Lymphoid enhancer-binding factor 1 antibody
    • T cell specific transcription factor 1 alpha antibody
    • T cell-specific transcription factor 1-alpha antibody
    • TCF 1 alpha antibody
    • TCF1 alpha antibody
    • TCF1-alpha antibody
    • TCF10 antibody
    • TCF1alpha antibody
    • TCF7L3 antibody
    • Transcription factor T cell specific 1 alpha antibody
    see all

Images

  • Immunohistochemical staining of paraffin embedded human tonsil with purified ab137872 at a working dilution of 1/500. The secondary antibody used is ab97051, an HRP-conjugated goat anti-rabbit IgG (H+L), at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  • Immunohistochemical staining of paraffin embedded rat spleen with purified ab137872 at a working dilution of 1/500. The secondary antibody used is ab97051, an HRP-conjugated goat anti-rabbit IgG (H+L), at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  • Immunohistochemical staining of paraffin-embedded human thymus with purified ab137872 at a working dilution of 1/500. The secondary antibody used is ab97051, an HRP-conjugated goat anti-rabbit IgG (H+L), at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  • ab137872 staining LEF1 in paraffin embedded mouse spleen tissue by Immunohistochemistry. Antigen retrieval was by heat mediation using ab93684 (Tris/EDTA buffer, ph 9).  Samples were incubated with primary antibody at 1:2000 dilution. A ready to use Goat Anti-rabbit IgG H&L (HRP) was used as the secondary antibody. Hematoxylin was used as a counter stain. Nuclear staining on T cells in periarterial lymphatic sheath of mouse spleen is observed (PMID: 21685909).

  • Anti-LEF1 antibody [EPR2029Y] (ab137872) at 1/2000 dilution (purified) + Rat thymus tissue lysate at 20 µg

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size: 44 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST

     

  • Anti-LEF1 antibody [EPR2029Y] (ab137872) at 1/10000 dilution (purified) + Jurkat (Human T cell leukemia cell line from peripheral blood) cell lysate at 10 µg

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size: 44 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST

  • Anti-LEF1 antibody [EPR2029Y] (ab137872) at 1/1000 dilution (purified) + Human fetal thymus lysate at 10 µg

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size: 44 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST

  • Anti-LEF1 antibody [EPR2029Y] (ab137872) at 1/1000 dilution + His-tagged mouse LEF-1 recombinant protein (aa1-397) at 0.01 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 44 kDa
    Observed band size: 44 kDa


    Exposure time: 1 second


    Blocking and diluting buffer: 5% NFDM/TBST

  • Immunofluorescence staining of Jurkat (Human T cell leukemia cell line from peripheral blood) cells with purified ab137872 at a working dilution of 1 in 500, counter-stained with DAPI. Tubulin was stained with mouse anti-tubulin at a dilution of 1/1000 (ab7291) and Alexa Fluor® 594 goat anti-mouse at a dilution of 1/500 (ab150120) . The secondary antibody was ab150077 Alexa Fluor® 488 goat anti rabbit, used at a dilution of 1 in 500. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100.

    The negative controls are shown in the bottom middle and right hand panels - for the first negative control, purified ab137872 was used at a dilution of 1/200 followed by an Alexa Fluor® 555 goat anti-mouse antibody at a dilution of 1/500 and for the second negative control mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab15007) were used.

  • Flow cytometric analysis of Jurkat cell line (human T cell leukemia T lymphocyte) fixed with 4% paraformaldehyde and permeabilized with 90% methanol labeling LEF1 with ab137872 at 1:600 dilution (red). This is compared with a Rabbit monoclonal IgG (ab172730) - Isotype control (black) and a unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti-rabbit IgG (Alexa Fluor® 488) was used as the secondary antibody.

  • Lane 1 (input): Rat thymus lysate, 10μg
    Lane 2 (+): Rat thymus lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of  ab137872 in rat thymus lysate

    ab137872 immunoprecipitating LEF1 in rat thymus lysate. For western blotting, primary antibody used was ab137872 at 1:1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

    Blocking and diluting buffer: 5% NFDM/TBST

    Exposure time: 3 minutes

  • Lane 1 (input): Jurkat (human T cell leukemia T lymphocyte) whole cell lysate, 10 μg
    Lane 2 (+): Jurkat whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of  ab137872 in Jurkat  whole cell lysate

    ab137872 immunoprecipitating LEF1 in Jurkat whole cell lysate. For western blotting, primary antibody used was ab137872 at 1:1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

    Blocking and diluting buffer: 5% NFDM/TBST

    Exposure time: 3 minutes

References

This product has been referenced in:
  • Ji Q  et al. Hematopoietic PBX-interacting protein mediates cartilage degeneration during the pathogenesis of osteoarthritis. Nat Commun 10:313 (2019). WB, IP ; Saccharomyces cerevisiae . Read more (PubMed: 30659184) »
  • Huang X  et al. Investigations on the mechanism of progesterone in inhibiting endometrial cancer cell cycle and viability via regulation of long noncoding RNA NEAT1/microRNA-146b-5p mediated Wnt/ß-catenin signaling. IUBMB Life 71:223-234 (2019). Read more (PubMed: 30452118) »
See all 19 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (skin)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: pH6 citrate
Permeabilization
No
Specification
skin
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Jun 18 2019

Application
IHC - Wholemount
Sample
Chicken Embryo (Whole embryo - stage HH9-10)
Specification
Whole embryo - stage HH9-10

Ana Azambuja

Verified customer

Submitted May 11 2018

Answer

Spleen should be a suitable positive control tissue for LEF1 detection.


For example, we have the following slides:


Spleen (human) normal tissue slides (ab4372)

Description: Spleen (human) normal tissue slides

Application: IHC-P

https://www.abcam.com/index.html?datasheet=4372 (or use the following: https://www.abcam.com/index.html?datasheet=4372).


.

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