RecombinantRabMAb

Recombinant Alexa Fluor® 488 Anti-LEF1 antibody [EPR2029Y] (ab246714)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Alexa Fluor® 488 Anti-LEF1 antibody [EPR2029Y] (ab246714)
  • Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 488 Anti-LEF1 antibody [EPR2029Y] (ab246714)
  • Flow Cytometry - Alexa Fluor® 488 Anti-LEF1 antibody [EPR2029Y] (ab246714)
  • Alexa Fluor® 488 Anti-LEF1 antibody [EPR2029Y] (ab246714)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Alexa Fluor® 488 Rabbit monoclonal [EPR2029Y] to LEF1
  • Suitable for: ICC/IF, Flow Cyt, IHC-P
  • Reacts with: Human
  • Conjugation: Alexa Fluor® 488. Ex: 495nm, Em: 519nm

Overview

  • Product name

    Alexa Fluor® 488 Anti-LEF1 antibody [EPR2029Y]
    See all LEF1 primary antibodies

  • Description

    Alexa Fluor® 488 Rabbit monoclonal [EPR2029Y] to LEF1

  • Host species

    Rabbit

  • Conjugation

    Alexa Fluor® 488. Ex: 495nm, Em: 519nm

  • Tested applications

    Suitable for: ICC/IF, Flow Cyt, IHC-Pmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

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

  • Positive control

    • ICC/IF: HT-29 cells. IHC-P: Normal human tonsil tissue sections.Flow Cyt: Jurkat cells.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

    Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@thermofisher.com.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Applications

Our Abpromise guarantee covers the use of ab246714 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
ICC/IF 1/100.

This product gave a positive signal in HT-29 fixed with 4% formaldehyde (10 min).
Flow Cyt 1/2500.
IHC-P 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Alexa Fluor® 488 Anti-LEF1 antibody [EPR2029Y] (ab246714)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Alexa Fluor® 488 Anti-LEF1 antibody [EPR2029Y] (ab246714)

      IHC image of LEF1 staining in a section of formalin-fixed paraffin-embedded normal human tonsil*.

      The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH9, epitope retrieval solution 2) for 20mins, performed on a Leica BOND™. Non-specific protein-protein interactions were then blocked in TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 1% (w/v) BSA for 1h at room temperature. The section was then incubated overnight at +4°C in TBS containing 0.025% (v/v) Triton X-100 and 1% (w/v) BSA with ab246714 at 1/100 dilution (shown in green) and counterstained using ab195884, Rat monoclonal to Tubulin (Alexa Fluor® 647), at 1/250 dilution (shown in red). Nuclear DNA was labeled with DAPI (shown in blue). The section was then mounted using Fluoromount®.

      Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

      For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

      *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

    • Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 488 Anti-LEF1 antibody [EPR2029Y] (ab246714)
      Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 488 Anti-LEF1 antibody [EPR2029Y] (ab246714)

      ab246714 staining LEF1 in HT-29 cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab246714 at 1/100 dilution (shown in green) and ab202272, Rabbit monoclonal to alpha Tubulin (Alexa Fluor® 594), at 1/250 dilution (shown in red). Nuclear DNA was labeled with DAPI (shown in blue).

      Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    • Flow Cytometry - Alexa Fluor® 488 Anti-LEF1 antibody [EPR2029Y] (ab246714)
      Flow Cytometry - Alexa Fluor® 488 Anti-LEF1 antibody [EPR2029Y] (ab246714)

      Overlay histogram showing Jurkat cells stained with ab246714 (red line). The cells were fixed with 4 % formaldehyde (10 min) and then permeabilized with 0.1 % PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction  followed by the antibody (ab246714) (1x 106 in 100µl at 0.2µg/ml (1/2500)) for 30 min at 22°C.

      Isotype control antibody (black line) was Rabbit IgG (monoclonal) Alexa Fluor 488® (ab199091) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

      Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.

    • Alexa Fluor® 488 Anti-LEF1 antibody [EPR2029Y] (ab246714)
      Alexa Fluor® 488 Anti-LEF1 antibody [EPR2029Y] (ab246714)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (0)

    Publishing research using ab246714? Please let us know so that we can cite the reference in this datasheet.

    ab246714 has not yet been referenced specifically in any publications.

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