Validated using a knockout cell lineRecombinantRabMAb

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

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

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: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free

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: Human
    Predicted to work with: Mouse, Rat

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: MCF7, 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.

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

The Abpromise guarantee

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).
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 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.

     

  • 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 MCF7 cell lysate
    Lane 2 : TLE1 CRISPR/Cas9 edited MCF7 cell lysate
    Lane 3 : SH-SY5Y cell lysate
    Lane 4 : 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 - 4: Merged signal (red and green). Green - ab183742 observed at 83 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

    ab183742 was shown to react with TLE 1 in western blot. The band observed in the CRISPR/Cas9 edited lysate lane below 83 kDa is likely to represent a truncated form. This has not been investigated further. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab183742 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h 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 : 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.

     

  • 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 (5)

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

ab183742 has been referenced in 5 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
  • Liu T  et al. lncRNA RASSF8-AS1 suppresses the progression of laryngeal squamous cell carcinoma via targeting the miR-664b-3p/TLE1 axis. Oncol Rep 44:2031-2044 (2020). PubMed: 33000257
  • 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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