Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-TLE 1 antibody [EPR9386(2)] (ab183742)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR9386(2)] to TLE 1
  • Suitable for: ICC/IF, IHC-P, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-TLE 1 antibody [EPR9386(2)]
    See all TLE 1 primary antibodies

  • Description

    Rabbit monoclonal [EPR9386(2)] to TLE 1

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC/IF, IHC-P, WBmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human TLE 1 aa 200-300. The exact sequence is proprietary.
    Database link: Q04724

  • Positive control

    • WB: HEK-293T, SH-SY5Y, HepG2, Jurkat and HeLa cell lysates. IHC-P: Human schwannoma and synovial sarcoma tissues. ICC/IF: MCF7 and HepG2 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.

    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

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.

  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 59% PBS, 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR9386(2)

  • Isotype

    IgG

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab183742 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.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB 1/1000 - 1/2000. Detects a band of approximately 83 kDa (predicted molecular weight: 83 kDa).

Target

  • Function

    Transcriptional corepressor that binds to a number of transcription factors. Inhibits NF-kappa-B-regulated gene expression. Inhibits the transcriptional activation mediated by FOXA2, and by CTNNB1 and TCF family members in Wnt signaling. The effects of full-length TLE family members may be modulated by association with dominant-negative AES. Unusual function as coactivator for ESRRG.

  • Tissue specificity

    In all tissues examined, mostly in brain, liver and muscle.

  • Sequence similarities

    Belongs to the WD repeat Groucho/TLE family.
    Contains 6 WD repeats.

  • Domain

    WD repeat Groucho/TLE family members are characterized by 5 regions, a glutamine-rich Q domain, a glycine/proline-rich GP domain, a central CcN domain, containing a nuclear localization signal, and a serine/proline-rich SP domain. The most highly conserved are the N-terminal Q domain and the C-terminal WD-repeat domain.

  • Post-translational
    modifications

    Phosphorylated, probably by CDK1. The degree of phosphorylation varies throughout the cell cycle, and is highest at the G2/M transition. Becomes hyperphosphorylated in response to cell differentiation and interaction with HES1 or RUNX1.
    Ubiquitinated by XIAP/BIRC4.

  • Cellular localization

    Nucleus. Nuclear and chromatin-associated, depending on isoforms and phosphorylation status. Hyperphosphorylation decreases the affinity for nuclear components.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • E(Sp1) homolog antibody
    • Enhancer of split groucho 1 antibody
    • Enhancer of split groucho like protein 1 antibody
    • Enhancer of split groucho-like protein 1 antibody
    • ESG antibody
    • ESG1 antibody
    • GRG1 antibody
    • TLE1 antibody
    • TLE1_HUMAN antibody
    • Transducin like enhancer of split 1 (E(sp1) homolog Drosophila) antibody
    • Transducin like enhancer of split 1 antibody
    • Transducin like enhancer of split 1 homolog of Drosophila E(sp1) antibody
    • Transducin like enhancer protein 1 antibody
    • Transducin-like enhancer protein 1 antibody
    see all

Images

  • Western blot - Anti-TLE 1 antibody [EPR9386(2)] (ab183742)
    Western blot - Anti-TLE 1 antibody [EPR9386(2)] (ab183742)
    All lanes : Anti-TLE 1 antibody [EPR9386(2)] (ab183742) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : TLE1 knockout HeLa cell lysate
    Lane 3 : 293T cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 83 kDa
    Observed band size: 83 kDa



    Lanes 1-3: Merged signal (red and green). Green - ab183742 observed at 83 kDa. Red - loading control, ab8245 observed at 37 kDa.

    ab183742 Anti-TLE 1 antibody [EPR9386(2)] was shown to specifically react with TLE 1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264901 (knockout cell lysate ab257241) was used. Wild-type and TLE 1 knockout samples were subjected to SDS-PAGE. ab183742 and Anti-GAPDH antibody [EPR16891] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

     

  • Immunocytochemistry/ Immunofluorescence - Anti-TLE 1 antibody [EPR9386(2)] (ab183742)
    Immunocytochemistry/ Immunofluorescence - Anti-TLE 1 antibody [EPR9386(2)] (ab183742)

    ab183742 staining TLE1 in wild-type MCF7 cells (top panel) and TLE1 knockout MCF7 cells (bottom panel). The cells were fixed with 4% paraformaldehyde (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 with ab183742 at 1/500 dilution and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 µg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 µg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
    Image was taken with a high-content analysis system (Perkin Elmer, Operetta CLS™).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TLE 1 antibody [EPR9386(2)] (ab183742)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TLE 1 antibody [EPR9386(2)] (ab183742)

    Immunohistochemical analysis of Human schwannoma, staining TLE 1 with ab183742 at 1/250 dilution. Detected using HRP Polymer for Rabbit IgG and counter-stained using hematoxylin.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Western blot - Anti-TLE 1 antibody [EPR9386(2)] (ab183742)
    Western blot - Anti-TLE 1 antibody [EPR9386(2)] (ab183742)
    All lanes : Anti-TLE 1 antibody [EPR9386(2)] (ab183742) at 1/1000 dilution

    Lane 1 : Wild-type HEK-293T cell lysate
    Lane 2 : TLE1 knockout HEK-293T cell lysate
    Lane 3 : HepG2 cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 83 kDa
    Observed band size: 83 kDa



    Lanes 1-3: Merged signal (red and green). Green - ab183742 observed at 83 kDa. Red - loading control, ab8245 observed at 37 kDa.

    ab183742 Anti-TLE 1 antibody [EPR9386(2)] was shown to specifically react with TLE 1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab265059 (knockout cell lysate ab257240) was used. Wild-type and TLE 1 knockout samples were subjected to SDS-PAGE. ab183742 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

     

  • Western blot - Anti-TLE 1 antibody [EPR9386(2)] (ab183742)
    Western blot - Anti-TLE 1 antibody [EPR9386(2)] (ab183742)
    All lanes : Anti-TLE 1 antibody [EPR9386(2)] (ab183742) at 1/1000 dilution

    Lane 1 : SH-SY5Y cell lysate
    Lane 2 : HepG2 cell lysate
    Lane 3 : Jurkat cell lysate
    Lane 4 : HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 83 kDa
    Observed band size: 83 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-TLE 1 antibody [EPR9386(2)] (ab183742)
    Immunocytochemistry/ Immunofluorescence - Anti-TLE 1 antibody [EPR9386(2)] (ab183742)

    Immunofluorescence analysis of paraformaldehyde-fixed HepG2 cells, staining TLE 1 (green) with ab183742 at 1/100 dilution. Alexa Fluor®488-conjugated goat anti rabbit IgG was used as a secondary antibody at 1/200 dilution. Nuclei were counterstained with DAPI (blue).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TLE 1 antibody [EPR9386(2)] (ab183742)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TLE 1 antibody [EPR9386(2)] (ab183742)

    Immunohistochemical analysis of Human synovial sarcoma, staining TLE 1 with ab183742 at 1/250 dilution. Detected using HRP Polymer for Rabbit IgG and counter-stained using hematoxylin.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Anti-TLE 1 antibody [EPR9386(2)] (ab183742)
    Anti-TLE 1 antibody [EPR9386(2)] (ab183742)

References (4)

Publishing research using ab183742? Please let us know so that we can cite the reference in this datasheet.

ab183742 has been referenced in 4 publications.

  • Wang Y  et al. Transducin-Like Enhancer of Split-1 Inhibits Malignant Behaviors in vitro and Predicts a Better Prognosis in Pancreatic Ductal Adenocarcinoma. Front Oncol 10:576 (2020). PubMed: 32432037
  • Takemoto J  et al. HuC/D expression in small round cell tumors and neuroendocrine tumors: a useful tool for distinguishing neuroblastoma from childhood small round cell tumors. Hum Pathol N/A:N/A (2018). PubMed: 30468801
  • Nowicki TS  et al. Infiltration of CD8 T Cells and Expression of PD-1 and PD-L1 in Synovial Sarcoma. Cancer Immunol Res 5:118-126 (2017). PubMed: 28039162
  • Piggin CL  et al. ELF5 isoform expression is tissue-specific and significantly altered in cancer. Breast Cancer Res 18:4 (2016). WB . PubMed: 26738740

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