Recombinant 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
Related conjugates and formulations
Overview
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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.
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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.
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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
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
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR9386(2) -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
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Related Products
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 |
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ICC/IF |
1/100.
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IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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WB |
1/1000 - 1/2000. Detects a band of approximately 83 kDa (predicted molecular weight: 83 kDa).
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Notes |
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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
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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
modificationsPhosphorylated, 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
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Database links
- Entrez Gene: 7088 Human
- Entrez Gene: 21885 Mouse
- Entrez Gene: 362533 Rat
- Omim: 600189 Human
- SwissProt: Q04724 Human
- SwissProt: Q62440 Mouse
- Unigene: 197320 Human
- Unigene: 689805 Human
see all -
Alternative names
- E(Sp1) homolog antibody
- Enhancer of split groucho 1 antibody
- Enhancer of split groucho like protein 1 antibody
see all
Images
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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 kDaLanes 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.
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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)
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.
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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 kDaLanes 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.
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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 kDaLanes 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.
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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 -
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).
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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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (7)
ab183742 has been referenced in 7 publications.
- Van Renterghem B et al. Enhanced Antitumor Efficacy of PhAc-ALGP-Dox, an Enzyme-Activated Doxorubicin Prodrug, in a Panel of THOP1-Expressing Patient-Derived Xenografts of Soft Tissue Sarcoma. Biomedicines 10:N/A (2022). PubMed: 35453612
- Bergeron BP et al. Epigenomic profiling of glucocorticoid responses identifies cis-regulatory disruptions impacting steroid resistance in childhood acute lymphoblastic leukemia. Leukemia 36:2374-2383 (2022). PubMed: 36028659
- 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