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TFE3 Recombinant Rabbit Monoclonal Antibody [JE60-60]

Usd: 385

Specification

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Catalog# HA721896

TFE3 Recombinant Rabbit Monoclonal Antibody [JE60-60]

Application

  • WB

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

TFE3 Recombinant Rabbit Monoclonal Antibody [JE60-60]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human TFE3 aa 150-499 / 575.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, FC

Target Molecular Weight

Predicted band size: 61 kDa

Positive Control

U-2 OS cell lysate, 293T cell lysate, C2C12 cell lysate, mouse testis tissue lysate, A-172 cell lysate, NIH/3T3 cell lysate, C6 cell lysate, PC-12 cell lysate, human testis tissue, human thyroid carcinoma tissue, rat testis tissue, U-2 OS.

Conjugation

unconjugated

Clone Number

JE60-60

RRID

AB_3096691

Product Features

Form

Liquid

Concentration

1 mg/ml(The concentration of this product may be batch-dependent)

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

  • 1:1,000

  • IHC-P

  • 1:800

  • FC

  • 1:1,000

Target

Function

Transcription factor that acts as a master regulator of lysosomal biogenesis and immune response. Specifically recognizes and binds E-box sequences (5'-CANNTG-3'); efficient DNA-binding requires dimerization with itself or with another MiT/TFE family member such as TFEB or MITF. Involved in the cellular response to amino acid availability by acting downstream of MTOR: in the presence of nutrients, TFE3 phosphorylation by MTOR promotes its cytosolic retention and subsequent inactivation. Upon starvation or lysosomal stress, inhibition of MTOR induces TFE3 dephosphorylation, resulting in nuclear localization and transcription factor activity. Maintains the pluripotent state of embryonic stem cells by promoting the expression of genes such as ESRRB; mTOR-dependent TFE3 cytosolic retention and inactivation promotes exit from pluripotency. Required to maintain the naive pluripotent state of hematopoietic stem cell; mTOR-dependent cytoplasmic retention of TFE3 promotes the exit of hematopoietic stem cell from pluripotency. Acts as a positive regulator of browning of adipose tissue by promoting expression of target genes; mTOR-dependent phosphorylation promotes cytoplasmic retention of TFE3 and inhibits browning of adipose tissue.

Background References

1. Lawrence R.E., Fromm S.A., Fu Y., Yokom A.L., Hurley J.H., Zoncu R. Structural mechanism of a Rag GTPase activation checkpoint by the lysosomal folliculin complex. Science 366:971-977 (2019)

2. Mathieu J., Detraux D., Kuppers D., Wang Y.,, Paddison P.J., Ruohola-Baker H. Folliculin regulates mTORC1/2 and WNT pathways in early human pluripotency. Nat. Commun. 10:632-632 (2019)

Subcellular Location

Cytoplasm, Nucleus.

UNIPROT

SwissProt: P19532 Human

SwissProt: Q64092 Mouse

Entrez Gene: 317376 Rat

Synonyms

bHLH e33 antibody

bHLHe33 antibody

Class E basic helix-loop-helix protein 33 antibody

RCCP 2 antibody

RCCP2 antibody

RCCX1 antibody

Renal cell carcinoma, papillary antibody

Tcfe 3 antibody

Tcfe3 antibody

TFE 3 antibody

Expand

Images

  • Western blot analysis of TFE3 on different lysates with Rabbit anti-TFE3 antibody (<a href="/products/HA721896" style="font-weight: bold;text-decoration: underline;">HA721896</a>) at 1/1,000 dilution.<br /><br />Lane 1: U-2 OS cell lysate (20 µg/Lane)<br />Lane 2: 293T cell lysate (20 µg/Lane)<br />Lane 3: C2C12 cell lysate (20 µg/Lane)<br />Lane 4: Mouse testis tissue lysate (40 µg/Lane)<br />Lane 5: A-172 cell lysate (30 µg/Lane)<br />Lane 6: NIH/3T3 cell lysate (30 µg/Lane)<br />Lane 7: C6 cell lysate (30 µg/Lane)<br />Lane 8: PC-12 cell lysate (30 µg/Lane)<br /><br />Predicted band size: 61 kDa<br />Observed band size: 70/80 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/HA721896" style="font-weight: bold;text-decoration: underline;">HA721896</a>) at 1/1,000 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.

    Western blot analysis of TFE3 on different lysates with Rabbit anti-TFE3 antibody (HA721896) at 1/1,000 dilution.

    Lane 1: U-2 OS cell lysate (20 µg/Lane)
    Lane 2: 293T cell lysate (20 µg/Lane)
    Lane 3: C2C12 cell lysate (20 µg/Lane)
    Lane 4: Mouse testis tissue lysate (40 µg/Lane)
    Lane 5: A-172 cell lysate (30 µg/Lane)
    Lane 6: NIH/3T3 cell lysate (30 µg/Lane)
    Lane 7: C6 cell lysate (30 µg/Lane)
    Lane 8: PC-12 cell lysate (30 µg/Lane)

    Predicted band size: 61 kDa
    Observed band size: 70/80 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 (HA721896) at 1/1,000 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.

  • Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-TFE3 antibody (<a href="/products/HA721896" style="font-weight: bold;text-decoration: underline;">HA721896</a>) at 1/800 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/HA721896" style="font-weight: bold;text-decoration: underline;">HA721896</a>) at 1/800 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 testis tissue with Rabbit anti-TFE3 antibody (HA721896) at 1/800 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 (HA721896) at 1/800 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 thyroid carcinoma tissue with Rabbit anti-TFE3 antibody (<a href="/products/HA721896" style="font-weight: bold;text-decoration: underline;">HA721896</a>) at 1/800 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/HA721896" style="font-weight: bold;text-decoration: underline;">HA721896</a>) at 1/800 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 thyroid carcinoma tissue with Rabbit anti-TFE3 antibody (HA721896) at 1/800 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 (HA721896) at 1/800 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 testis tissue with Rabbit anti-TFE3 antibody (<a href="/products/HA721896" style="font-weight: bold;text-decoration: underline;">HA721896</a>) at 1/800 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/HA721896" style="font-weight: bold;text-decoration: underline;">HA721896</a>) at 1/800 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 testis tissue with Rabbit anti-TFE3 antibody (HA721896) at 1/800 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 (HA721896) at 1/800 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.

  • Flow cytometric analysis of U-2 OS cells labeling TFE3.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA721896" style="font-weight: bold;text-decoration: underline;">HA721896</a>, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of U-2 OS cells labeling TFE3.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA721896, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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