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LEF1 Recombinant Rabbit Monoclonal Antibody [JE52-28]

Usd: 385

Specification

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Catalog# HA721992

LEF1 Recombinant Rabbit Monoclonal Antibody [JE52-28]

Application

  • WB

  • IF-Cell

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

LEF1 Recombinant Rabbit Monoclonal Antibody [JE52-28]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human LEF1 aa 121-170 / 399.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P

Molecular Weight

Predicted band size: 44 kDa

Positive Control

Jurkat cell lysate, mouse thymus tissue lysate, rat thymus tissue lysate, Jurkat, human lymph node tissue, human thymus tissue, human tonsil tissue, rat thymus tissue.

Conjugation

unconjugated

Clone Number

JE52-28

RRID

AB_3096797

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000

  • IF-Cell

  • 1:500

  • IHC-P

  • 1:500-1:2,000

Target

Function

Lymphoid enhancer-binding factor 1 (LEF1) is a protein that in humans is encoded by the LEF1 gene. It's a member of T cell factor/lymphoid enhancer factor (TCF/LEF) family. Lymphoid enhancer-binding factor-1 (LEF1) is a 48-kD nuclear protein that is expressed in pre-B and T cells. It binds to a functionally important site in the T-cell receptor-alpha (TCRA) enhancer and confers maximal enhancer activity. LEF1 belongs to a family of regulatory proteins that share homology with high mobility group protein-1 (HMG1). LEF1 is highly overexpressed and associated with disease progression and poor prognosis in B-cell chronic lymphocytic leukemia and other kinds of malignancies like colorectal cancer. It is also a promising potential drug target.

Background References

1. Fakhr E et al. LEF1 silencing sensitizes colorectal cancer cells to oxaliplatin, 5-FU, and irinotecan. Biomed Pharmacother. 2021 Nov

2. Zhang Z et al. Asporin represses gastric cancer apoptosis via activating LEF1-mediated gene transcription independent of β-catenin. Oncogene. 2021 Jul

Subcellular Location

Nucleus.

UNIPROT

SwissProt: Q9UJU2 Human

SwissProt: P27782 Mouse

SwissProt: Q9QXN1 Rat

Synonyms

DKFZp586H0919 antibody

FLJ46390 antibody

LEF 1 antibody

LEF-1 antibody

Lef1 antibody

LEF1_HUMAN antibody

Lymphoid enhancer binding factor 1 antibody

Lymphoid enhancer-binding factor 1 antibody

T cell specific transcription factor 1 alpha antibody

T cell-specific transcription factor 1-alpha antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of LEF1 on different lysates with Rabbit anti-LEF1 antibody (<a href="/products/HA721992" style="font-weight: bold;text-decoration: underline;">HA721992</a>) at 1/2,000 dilution.<br /><br />Lane 1: Jurkat cell lysate (20 µg/Lane)<br />Lane 2: HeLa cell lysate (negative) (20 µg/Lane)<br />Lane 3: Mouse thymus tissue lysate (40 µg/Lane)<br />Lane 4: Rat thymus tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 44 kDa<br />Observed band size: 35-55 kDa<br /><br />Exposure time: 3 minutes;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA721992" style="font-weight: bold;text-decoration: underline;">HA721992</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Relative expression (RE)

    Western blot analysis of LEF1 on different lysates with Rabbit anti-LEF1 antibody (HA721992) at 1/2,000 dilution.

    Lane 1: Jurkat cell lysate (20 µg/Lane)
    Lane 2: HeLa cell lysate (negative) (20 µg/Lane)
    Lane 3: Mouse thymus tissue lysate (40 µg/Lane)
    Lane 4: Rat thymus tissue lysate (40 µg/Lane)

    Predicted band size: 44 kDa
    Observed band size: 35-55 kDa

    Exposure time: 3 minutes;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721992) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of LEF1 on different lysates with Rabbit anti-LEF1 antibody (<a href="/products/HA721992" style="font-weight: bold;text-decoration: underline;">HA721992</a>) at 1/2,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate (10 µg/Lane)<br />Lane 2: HAP1-LEF1 KD cell lysate (10 µg/Lane)<br /><br />Predicted band size: 44 kDa<br />Observed band size: 35-55 kDa<br /><br />Exposure time: 3 minutes;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA721992" style="font-weight: bold;text-decoration: underline;">HA721992</a>) at 1/2,000 dilution was used in <a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a> at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of LEF1 on different lysates with Rabbit anti-LEF1 antibody (HA721992) at 1/2,000 dilution.

    Lane 1: HAP1-parental cell lysate (10 µg/Lane)
    Lane 2: HAP1-LEF1 KD cell lysate (10 µg/Lane)

    Predicted band size: 44 kDa
    Observed band size: 35-55 kDa

    Exposure time: 3 minutes;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721992) at 1/2,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Immunocytochemistry analysis of Jurkat (positive) and HeLa (negative) labeling LEF1 with Rabbit anti-LEF1 antibody (<a href="/products/HA721992" style="font-weight: bold;text-decoration: underline;">HA721992</a>) at 1/500 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-LEF1 antibody (<a href="/products/HA721992" style="font-weight: bold;text-decoration: underline;">HA721992</a>) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    ☑ Relative expression (RE)

    Immunocytochemistry analysis of Jurkat (positive) and HeLa (negative) labeling LEF1 with Rabbit anti-LEF1 antibody (HA721992) at 1/500 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-LEF1 antibody (HA721992) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Immunohistochemical analysis of paraffin-embedded human lymph node tissue with Rabbit anti-LEF1 antibody (<a href="/products/HA721992" style="font-weight: bold;text-decoration: underline;">HA721992</a>) at 1/2,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA721992" style="font-weight: bold;text-decoration: underline;">HA721992</a>) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human lymph node tissue with Rabbit anti-LEF1 antibody (HA721992) at 1/2,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721992) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human thymus tissue with Rabbit anti-LEF1 antibody (<a href="/products/HA721992" style="font-weight: bold;text-decoration: underline;">HA721992</a>) at 1/2,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA721992" style="font-weight: bold;text-decoration: underline;">HA721992</a>) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human thymus tissue with Rabbit anti-LEF1 antibody (HA721992) at 1/2,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721992) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-LEF1 antibody (<a href="/products/HA721992" style="font-weight: bold;text-decoration: underline;">HA721992</a>) at 1/500 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA721992" style="font-weight: bold;text-decoration: underline;">HA721992</a>) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-LEF1 antibody (HA721992) at 1/500 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721992) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded rat thymus tissue with Rabbit anti-LEF1 antibody (<a href="/products/HA721992" style="font-weight: bold;text-decoration: underline;">HA721992</a>) at 1/2,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA721992" style="font-weight: bold;text-decoration: underline;">HA721992</a>) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded rat thymus tissue with Rabbit anti-LEF1 antibody (HA721992) at 1/2,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721992) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

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