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mTOR Recombinant Rabbit Monoclonal Antibody [SU30-00]

Usd: 385

Specification

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Catalog# ET1608-5

mTOR Recombinant Rabbit Monoclonal Antibody [SU30-00]

Application

  • WB

  • IHC-P

  • IP

  • IF-Cell

  • IF-Tissue

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

mTOR Recombinant Rabbit Monoclonal Antibody [SU30-00]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within human mTOR aa 2400-2440.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, IP, IF-Cell, IF-Tissue

Molecular Weight

Predicted band size: 289 kDa

Positive Control

HeLa cell lysate, MCF7 cell lysate, HEK-293 cell lysate, HepG2 cell lysate, C2C12 cell lysate, PC-12 cell lysate, C6 cell lysate, mouse testis tissue lysate, rat testis tissue lysate, rat heart tissue lysate, SiHa cell lysate, human breast carcinoma tissue, HeLa, NIH/3T3, human kidney tissue, mouse testis tissue, mouse kidney tissue.

Conjugation

unconjugated

Clone Number

SU30-00

RRID

AB_2928996

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

  • 1:5,000

  • IHC-P

  • 1:200-1:1,000

  • IF-cell

  • 1:50-1:200

  • IP

  • Use at an assay dependent concentration.

  • IF-Tissue

  • 1:500

Target

Function

The mammalian target of rapamycin (mTOR), also referred to as the mechanistic target of rapamycin, and sometimes called FK506-binding protein 12-rapamycin-associated protein 1 (FRAP1), is a kinase that in humans is encoded by the MTOR gene. mTOR is a member of the phosphatidylinositol 3-kinase-related kinase family of protein kinases. mTOR links with other proteins and serves as a core component of two distinct protein complexes, mTOR complex 1 and mTOR complex 2, which regulate different cellular processes. In particular, as a core component of both complexes, mTOR functions as a serine/threonine protein kinase that regulates cell growth, cell proliferation, cell motility, cell survival, protein synthesis, autophagy, and transcription. As a core component of mTORC2, mTOR also functions as a tyrosine protein kinase that promotes the activation of insulin receptors and insulin-like growth factor 1 receptors. mTORC2 has also been implicated in the control and maintenance of the actin cytoskeleton.

Background References

1. Atif F et al. Anti-tumor effects of progesterone in human glioblastoma multiforme: role of PI3K/Akt/mTOR signaling. J Steroid Biochem Mol Biol 146:62-73 (2015).

2. Atif F et al. The Synergistic Effect of Combination Progesterone and Temozolomide on Human Glioblastoma Cells. PLoS One 10:e0131441 (2015).

Sequence Similarity

Belongs to the PI3/PI4-kinase family.

Tissue Specificity

Expressed in numerous tissues, with highest levels in testis.

Post-translational Modification

Autophosphorylates when part of mTORC1 or mTORC2. Phosphorylation at Ser-1261, Ser-2159 and Thr-2164 promotes autophosphorylation. Phosphorylation in the kinase domain modulates the interactions of MTOR with RPTOR and PRAS40 and leads to increased intrinsic mTORC1 kinase activity. Phosphorylation at Thr-2173 in the ATP-binding region by AKT1 strongly reduces kinase activity.

Subcellular Location

Endoplasmic reticulum membrane, Microsome membrane, PML body, Golgi apparatus membrane, Cytoplasm, Lysosome, Lysosome membrane, Mitochondrion outer membrane, phagosome.

UNIPROT

SwissProt: P42345 Human

SwissProt: Q9JLN9 Mouse

SwissProt: P42346 Rat

Synonyms

dJ576K7.1 (FK506 binding protein 12 rapamycin associated protein 1) antibody

FK506 binding protein 12 rapamycin associated protein 1 antibody

FK506 binding protein 12 rapamycin associated protein 2 antibody

FK506 binding protein 12 rapamycin complex associated protein 1 antibody

FK506-binding protein 12-rapamycin complex-associated protein 1 antibody

FKBP rapamycin associated protein antibody

FKBP12 rapamycin complex associated protein antibody

FKBP12-rapamycin complex-associated protein 1 antibody

FKBP12-rapamycin complex-associated protein antibody

FLJ44809 antibody

Expand

Images

  • Western blot analysis of mTOR on different lysates with Rabbit anti-mTOR antibody (<a href="/products/ET1608-5" style="font-weight: bold;text-decoration: underline;">ET1608-5</a>) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate (20 µg/Lane)<br />Lane 2: MCF7 cell lysate (20 µg/Lane)<br />Lane 3: HEK-293 cell lysate (20 µg/Lane)<br />Lane 4: HepG2 cell lysate (20 µg/Lane)<br />Lane 5: C2C12 cell lysate (20 µg/Lane)<br />Lane 6: PC-12 cell lysate (20 µg/Lane)<br />Lane 7: C6 cell lysate (20 µg/Lane)<br />Lane 8: Mouse testis tissue lysate (20 µg/Lane)<br />Lane 9: Rat testis tissue lysate (20 µg/Lane)<br /><br />Predicted band size: 289 kDa<br />Observed band size: 289 kDa<br /><br />Exposure time: 24 seconds; ECL: K1801;<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/ET1608-5" style="font-weight: bold;text-decoration: underline;">ET1608-5</a>) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution were 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 mTOR on different lysates with Rabbit anti-mTOR antibody (ET1608-5) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution.

    Lane 1: HeLa cell lysate (20 µg/Lane)
    Lane 2: MCF7 cell lysate (20 µg/Lane)
    Lane 3: HEK-293 cell lysate (20 µg/Lane)
    Lane 4: HepG2 cell lysate (20 µg/Lane)
    Lane 5: C2C12 cell lysate (20 µg/Lane)
    Lane 6: PC-12 cell lysate (20 µg/Lane)
    Lane 7: C6 cell lysate (20 µg/Lane)
    Lane 8: Mouse testis tissue lysate (20 µg/Lane)
    Lane 9: Rat testis tissue lysate (20 µg/Lane)

    Predicted band size: 289 kDa
    Observed band size: 289 kDa

    Exposure time: 24 seconds; ECL: K1801;
    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 (ET1608-5) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution were 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.

  • Immunocytochemistry analysis of HeLa cells labeling mTOR with Rabbit anti-mTOR antibody (<a href="/products/ET1608-5" style="font-weight: bold;text-decoration: underline;">ET1608-5</a>) at 1/100 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-mTOR antibody (<a href="/products/ET1608-5" style="font-weight: bold;text-decoration: underline;">ET1608-5</a>) at 1/100 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.

    Immunocytochemistry analysis of HeLa cells labeling mTOR with Rabbit anti-mTOR antibody (ET1608-5) at 1/100 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-mTOR antibody (ET1608-5) at 1/100 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.

  • Immunocytochemistry analysis of NIH/3T3 cells labeling mTOR with Rabbit anti-mTOR antibody (<a href="/products/ET1608-5" style="font-weight: bold;text-decoration: underline;">ET1608-5</a>) at 1/200 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-mTOR antibody (<a href="/products/ET1608-5" style="font-weight: bold;text-decoration: underline;">ET1608-5</a>) at 1/200 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.

    Immunocytochemistry analysis of NIH/3T3 cells labeling mTOR with Rabbit anti-mTOR antibody (ET1608-5) at 1/200 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-mTOR antibody (ET1608-5) at 1/200 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.

  • Immunocytochemistry analysis of PC-12 cells labeling mTOR with Rabbit anti-mTOR antibody (<a href="/products/ET1608-5" style="font-weight: bold;text-decoration: underline;">ET1608-5</a>) at 1/100 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-mTOR antibody (<a href="/products/ET1608-5" style="font-weight: bold;text-decoration: underline;">ET1608-5</a>) at 1/100 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.

    Immunocytochemistry analysis of PC-12 cells labeling mTOR with Rabbit anti-mTOR antibody (ET1608-5) at 1/100 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-mTOR antibody (ET1608-5) at 1/100 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.

  • Application: IF-Tissue<br /><br />Species: Mouse<br /><br />Site: testis<br /><br />Sample: Paraffin-embedded section<br /><br />Antibody concentration: 1/500

    Application: IF-Tissue

    Species: Mouse

    Site: testis

    Sample: Paraffin-embedded section

    Antibody concentration: 1/500

  • Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-mTOR antibody (<a href="/products/ET1608-5" style="font-weight: bold;text-decoration: underline;">ET1608-5</a>) at 1/5,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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/ET1608-5" style="font-weight: bold;text-decoration: underline;">ET1608-5</a>) at 1/5,000 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 kidney tissue with Rabbit anti-mTOR antibody (ET1608-5) at 1/5,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (ET1608-5) at 1/5,000 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 testis tissue with Rabbit anti-mTOR antibody (<a href="/products/ET1608-5" style="font-weight: bold;text-decoration: underline;">ET1608-5</a>) at 1/5,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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/ET1608-5" style="font-weight: bold;text-decoration: underline;">ET1608-5</a>) at 1/5,000 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 testis tissue with Rabbit anti-mTOR antibody (ET1608-5) at 1/5,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (ET1608-5) at 1/5,000 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.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

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