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Low endotoxin, Azide free.
Our first-to-market chimera with mouse IgG2c backbone, this functional antibody specifically depletes neutrophils in vivo for up to 72h.
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Rabbit anti-LEF1 antibody EPR2029Y ab137872 is a rabbit monoclonal antibody that is used in LEF1 western blotting, IHC, immunofluorescence and flow cytometry. Suitable for human, mouse and rat samples.
Recombinant format for high batch-to-batch consistency and reproducible results
Specificity confirmed with Lef1 knockout cell line validation
Specificity and sensitivity confirmed in IHC with multi-tissue microarray validation.
Anti-LEF1 antibody clone EPR2029Y is cited in over 80 publications
IgG
Rabbit
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
IHC-P | IP | WB | ICC/IF | Flow Cyt (Intra) | |
---|---|---|---|---|---|
Human | Tested | Tested | Tested | Tested | Tested |
Mouse | Tested | Expected | Expected | Expected | Expected |
Rat | Tested | Tested | Tested | Expected | Expected |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/100 - 1/500 | Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Species Rat | Dilution info 1/100 - 1/500 | Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Species Human | Dilution info 1/100 - 1/500 | Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Rat, Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info 1/1000 | Notes We don't recommend this antibody for mouse in Western Blot. In our hands an extra band was observed in mouse tissue lysates. |
Species Human | Dilution info 1/1000 | Notes We don't recommend this antibody for mouse in Western Blot. In our hands an extra band was observed in mouse tissue lysates. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/500 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Transcription factor that binds DNA in a sequence-specific manner (PubMed:2010090). Participates in the Wnt signaling pathway (By similarity). Activates transcription of target genes in the presence of CTNNB1 and EP300 (By similarity). PIAG antagonizes both Wnt-dependent and Wnt-independent activation by LEF1 (By similarity). TLE1, TLE2, TLE3 and TLE4 repress transactivation mediated by LEF1 and CTNNB1 (PubMed:11266540). Regulates T-cell receptor alpha enhancer function (PubMed:19653274). Required for IL17A expressing gamma-delta T-cell maturation and development, via binding to regulator loci of BLK to modulate expression (By similarity). May play a role in hair cell differentiation and follicle morphogenesis (By similarity).Isoform 1Transcriptionally activates MYC and CCND1 expression and enhances proliferation of pancreatic tumor cells.Isoform 3Lacks the CTNNB1 interaction domain and may therefore be an antagonist for Wnt signaling.Isoform 5Transcriptionally 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.
Lymphoid enhancer-binding factor 1, LEF-1, T cell-specific transcription factor 1-alpha, TCF1-alpha, LEF1
Rabbit anti-LEF1 antibody EPR2029Y ab137872 is a rabbit monoclonal antibody that is used in LEF1 western blotting, IHC, immunofluorescence and flow cytometry. Suitable for human, mouse and rat samples.
Recombinant format for high batch-to-batch consistency and reproducible results
Specificity confirmed with Lef1 knockout cell line validation
Specificity and sensitivity confirmed in IHC with multi-tissue microarray validation.
Anti-LEF1 antibody clone EPR2029Y is cited in over 80 publications
Lymphoid enhancer-binding factor 1, LEF-1, T cell-specific transcription factor 1-alpha, TCF1-alpha, LEF1
IgG
Rabbit
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
EPR2029Y
Affinity purification Protein A
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
LEF1 plays a significant role in the regulation of cell fate decisions and development. It acts as part of a transcription complex that binds to specific DNA sequences facilitating the transcription of target genes. LEF1 interacts with ?-catenin and this interaction is essential for the regulation of gene expression during embryonic development and cellular proliferation. LEF1's biological functions make it a useful marker in various research applications and assays like LEF1 ELISA are often utilized to study its expression levels in different physiological contexts.
LEF1 (Lymphoid Enhancer-Binding Factor 1) also known as LEF-1 or LEF 1 functions as a transcription factor involved in Wnt signaling pathways. The LEF1 protein has a mass of about 45 kilodaltons. Expression of LEF1 occurs mainly in lymphoid tissues and the central nervous system. Researchers often use LEF1 in immunohistochemistry (LEF1 IHC) for its ability to bind DNA at specific sites regulating gene expression important for cellular development and differentiation especially in T cells such as Jurkat cells.
LEF1 is a central player in the canonical Wnt/?-catenin signaling pathway. This pathway is fundamental for processes like embryogenesis and cellular growth. LEF1 interacts closely with proteins such as TCF (T-cell factor) family members and ?-catenin to regulate gene expression. Disruption in the Wnt pathway where LEF1 is significant can lead to abnormal cell growth and differentiation. Consequently LEF1 serves as an indicator and potential regulator of pathway activities in various cellular environments.
LEF1 is associated with numerous conditions particularly in oncology and immunology. Aberrations in LEF1 expression contribute to diseases like leukemia and lymphoma. Overexpression or mutations of LEF1 might lead to unregulated Wnt signaling promoting cancer progression. Furthermore LEF1's interactions with proteins such as MYC a well-known oncogene highlight its importance in tumorigenesis. Understanding LEF1's mechanisms can offer insights into therapeutic targets for these diseases providing a pathway for research into drug development and intervention strategies.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
Lane 1 (input): Jurkat (human T cell leukemia T lymphocyte) whole cell lysate, 10 μg
Lane 2 (+): Jurkat whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab137872 in Jurkat whole cell lysate
ab137872 immunoprecipitating LEF1 in Jurkat whole cell lysate. For western blotting, primary antibody used was ab137872 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST
Exposure time: 3 minutes
All lanes: Immunoprecipitation - Anti-LEF1 antibody [EPR2029Y] (AB137872)
Predicted band size: 44 kDa
Exposure time: 3min
Blocking/Dilution buffer: 5% NFDM/TBST
All lanes: Western blot - Anti-LEF1 antibody [EPR2029Y] (AB137872) at 1/1000 dilution
All lanes: Human fetal thymus lysate at 10 µg
All lanes: HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 44 kDa
Immunohistochemical staining of paraffin embedded human tonsil with purified ab137872 at a working dilution of 1/500. The secondary antibody used is ab97051, an HRP-conjugated goat anti-rabbit IgG (H+L), at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
False colour image of Western blot: Anti-LEF1 antibody [EPR2029Y] staining at 1/1000 dilution, shown in black; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab137872 was shown to bind specifically to LEF1. A band was observed at 40/53 kDa in wild-type Jurkat cell lysates with no signal observed at this size in Lef1 knockout cell line ab274898 (knockout cell lysate ab274956). To generate this image, wild-type and Lef1 knockout Jurkat cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times before development with Optiblot (ECL reagent ab133456) and imaged with 4 minutes exposure time. Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-LEF1 antibody [EPR2029Y] (AB137872) at 1/1000 dilution
Lane 1: Wild-type Jurkat cell lysate at 40 µg
Lane 2: Lef1 knockout Jurkat cell lysate at 40 µg
Lane 3: Jurkat cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 44 kDa
Observed band size: 40 kDa
Immunofluorescence staining of Jurkat (Human T cell leukemia cell line from peripheral blood) cells with purified ab137872 at a working dilution of 1 in 500, counter-stained with DAPI. Tubulin was stained with mouse anti-tubulin at a dilution of 1/1000 (ab7291) and Alexa Fluor® 594 goat anti-mouse at a dilution of 1/500 (ab150120) . The secondary antibody was ab150077 Alexa Fluor® 488 goat anti rabbit, used at a dilution of 1 in 500. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100.
The negative controls are shown in the bottom middle and right hand panels - for the first negative control, purified ab137872 was used at a dilution of 1/200 followed by an Alexa Fluor® 555 goat anti-mouse antibody at a dilution of 1/500 and for the second negative control mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab15007) were used.
Intracellular flow cytometric analysis of Jurkat cell line (human T cell leukemia T lymphocyte) fixed with 4% paraformaldehyde and permeabilized with 90% methanol labeling LEF1 with ab137872 at 1/600 dilution (red). This is compared with a Rabbit monoclonal IgG (ab172730) - Isotype control (black) and a unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti-rabbit IgG (Alexa Fluor®488) was used as the secondary antibody.
Different batches of ab137872 were tested on Jurkat (Human T cell leukemia T lymphocyte) lysate at 1.1 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 25-57 kDa.
All lanes: Western blot - Anti-LEF1 antibody [EPR2029Y] (AB137872)
Predicted band size: 44 kDa
Immunohistochemical staining of paraffin embedded rat spleen with purified ab137872 at a working dilution of 1/500. The secondary antibody used is ab97051, an HRP-conjugated goat anti-rabbit IgG (H+L), at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Immunohistochemical staining of paraffin-embedded human thymus with purified ab137872 at a working dilution of 1/500. The secondary antibody used is ab97051, an HRP-conjugated goat anti-rabbit IgG (H+L), at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Lane 1 (input): Rat thymus lysate, 10μg
Lane 2 (+): Rat thymus lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab137872 in rat thymus lysate
ab137872 immunoprecipitating LEF1 in rat thymus lysate. For western blotting, primary antibody used was ab137872 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST
Exposure time: 3 minutes
All lanes: Immunoprecipitation - Anti-LEF1 antibody [EPR2029Y] (AB137872)
Predicted band size: 44 kDa
Exposure time: 3min
ab137872 staining LEF1 in paraffin embedded mouse spleen tissue by Immunohistochemistry. Antigen retrieval was performed by heat mediation using ab93684 (Tris/EDTA buffer, ph 9). Samples were incubated with primary antibody at 1/2000 dilution. A ready to use Goat Anti-rabbit IgG H&L (HRP) was used as the secondary antibody. Hematoxylin was used as a counterstain. Nuclear staining on T cells in periarterial lymphatic sheath of mouse spleen is observed (PMID: 21685909).
Blocking/Dilution buffer: 5% NFDM/TBST
All lanes: Western blot - Anti-LEF1 antibody [EPR2029Y] (AB137872) at 1/2000 dilution
All lanes: Rat thymus tissue lysate at 20 µg
All lanes: HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 44 kDa
Blocking/Dilution buffer: 5% NFDM/TBST
All lanes: Western blot - Anti-LEF1 antibody [EPR2029Y] (AB137872) at 1/10000 dilution
All lanes: Jurkat (Human T cell leukemia cell line from peripheral blood) cell lysate at 10 µg
All lanes: HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 44 kDa
Blocking and diluting buffer: 5% NFDM/TBST
All lanes: Western blot - Anti-LEF1 antibody [EPR2029Y] (AB137872) at 1/1000 dilution
All lanes: His-tagged mouse LEF-1 recombinant protein (aa1-397) at 0.01 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051) at 1/100000 dilution
Predicted band size: 44 kDa
Observed band size: 44 kDa
Exposure time: 1s
Tissue Microarrays stained for Anti-LEF1 antibody [EPR2029Y] using ab137872 in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negaive (cross mark) staining per sample type tested. The sections were incubated with ab137872 for 30 mins at room temperature used at 1:2000 dilution (1.05 μg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB secondary antibody (ab209101). Counterstain was Hematoxylin.
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
ab137872 staining LEF1 in paraffin embedded human melanoma tissue by Immunohistochemistry. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins. Samples were incubated with primary antibody at 1/2000 dilution for 30 mins at room temperature. A ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Hematoxylin was used as a counterstain. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
ab137872 staining LEF1 in paraffin embedded human thymona tissue by Immunohistochemistry. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins. Samples were incubated with primary antibody at 1/2000 dilution for 30 mins at room temperature. A ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Hematoxylin was used as a counterstain. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Immunohistochemical analysis of formalin fixed paraffin embedded human tonsil labelling LEF1 with ab137872 at a concentration of 0.5 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32 mins at 100°C with ULTRA cell conditioning solution (CC1, pH 8.5). ab137872 anti LEF1 antibody [EPR2029Y] was incubated at 37°C for 16 mins. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.
Flow cytometry overlay histogram showing left Jurkat positive cells and right negative HeLa stained with ab137872 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilised 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 (ab137872) (1x 106 in 100μl at 0.2μg/ml (1/11500)) for 30min at 22°C.
The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C
Isotype control antibody (black line) was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
This antibody gave a positive signal in Jurkat Fixed with 4% formaldehyde (10 min) / permeabilised with 0.1% PBS-Triton X-100 for 15 min under the same conditions.
ab137872 staining LEF1 in paraffin embedded human melanoma tissue by Immunohistochemistry. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins. Samples were incubated with primary antibody at 1/2000 dilution for 30 mins at room temperature. A ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Hematoxylin was used as a counterstain. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
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