Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP2030Y] to LEF1
- Suitable for: ICC/IF, WB, Flow Cyt, IHC-P
- Reacts with: Human
Product nameAnti-LEF1 antibody [EP2030Y]
See all LEF1 primary antibodies
DescriptionRabbit monoclonal [EP2030Y] to LEF1
Tested applicationsSuitable for: ICC/IF, WB, Flow Cyt, IHC-Pmore details
Species reactivityReacts with: Human
corresponding to Human LEF1 aa 250-350 (internal sequence).
- Purchase matching WB positive control:Recombinant Human LEF1 protein
- ICC/IF: Ramos and Jurkat cells. IHC-P: Human colon adenocarcinoma, human tonsil tissue. Flow Cyt: Jurkat cells. WB: Jurkat cell lysate
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Storage instructionsShipped 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 bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
PurityProtein A purified
Our Abpromise guarantee covers the use of ab53293 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||1/50 - 1/250.|
|WB||1/5000. Detects a band of approximately 55 kDa (predicted molecular weight: 44 kDa).
For unpurified format use at 1/1000 dilution.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
|IHC-P||1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.|
FunctionParticipates 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 specificityDetected 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 similaritiesBelongs to the TCF/LEF family.
Contains 1 HMG box DNA-binding domain.
DomainProline-rich and acidic regions are implicated in the activation functions of RNA polymerase II transcription factors.
Cellular localizationNucleus. Found in nuclear bodies upon PIASG binding.
- Information by UniProt
- DKFZp586H0919 antibody
- FLJ46390 antibody
- LEF 1 antibody
Anti-LEF1 antibody [EP2030Y] (ab53293) at 1/5000 dilution + Jurkat (Human T cell leukemia T lymphocyte) whole cell lysates at 15 µg
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 44 kDa
Observed band size: 23-44 kDa why is the actual band size different from the predicted?
This antibody recognizes all 7 LEF1 isoforms with the MW ranging from 23 to 44 KD based on immunogen blast.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human tonsil tissue sections labeling LEF1 with purified ab53293 at 1/50 dilution (1.94 µg/ml). Heat mediated antigen retrieval was performed using heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
Immunocytochemistry/ Immunofluorescence analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling LEF1 with purified ab53293 at 1/50 dilution (2.0 µg/ml). Cells were fixed in 100% Methanol. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Flow Cytometry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling LEF1 with purified ab53293 at 1/20 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
ab53293 (unpurified) staining LEF1 in RAMOS (human Burkitt's lymphoma) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/500. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a dilution of 1/1000. DAPI was used as a nuclear counterstain and the negative control was PBS only.
Paraffin-embedded human colon adenocarcinoma tissue stained for LEF1 using ab53293 (unpurified) at 1/50 dilution in immunohistochemical analysis.
Flow cytometric analysis of permeabilized Jurkat cells using ab53293 (unpurified) (red) or a rabbit IgG (negative) (green).
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.
ab53293 has been referenced in 12 publications.
- Tang L et al. Long Noncoding RNA DCST1-AS1 Promotes Cell Proliferation and Metastasis in Triple-negative Breast Cancer by Forming a Positive Regulatory Loop with miR-873-5p and MYC. J Cancer 11:311-323 (2020). PubMed: 31897227
- Hitomi Y et al. NFKB1 and MANBA Confer Disease Susceptibility to Primary Biliary Cholangitis via Independent Putative Primary Functional Variants. Cell Mol Gastroenterol Hepatol 7:515-532 (2019). PubMed: 30528300
- Zhang XF et al. LEF-1 gene silencing inhibits pulmonary vascular remodeling and occurrence of pulmonary arterial hypertension through the ß-catenin signaling pathway. Biomed Pharmacother 108:817-827 (2018). PubMed: 30372893
- Qiu J et al. MiR-557 works as a tumor suppressor in human lung cancers by negatively regulating LEF1 expression. Tumour Biol 39:1010428317709467 (2017). PubMed: 28639890
- Murakami T et al. Isolation and characterization of lymphoid enhancer factor-1-positive deciduous dental pulp stem-like cells after transfection with a piggyBac vector containing LEF1 promoter-driven selection markers. Arch Oral Biol 81:110-120 (2017). ICC/IF ; Human . PubMed: 28500952