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Phospho-mTOR (S2448) Mouse Monoclonal Antibody [A5D5]

Usd: 350

Specification

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Catalog# HA600094

Phospho-mTOR (S2448) Mouse Monoclonal Antibody [A5D5]

Application

  • WB

  • IHC-P

Reactivity

  • Human

  • Mouse

Predicted reactivity

  • Rat

Predicted species support after-sales service
Conjugation

  • unconjugated

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

Overview

Product Name

Phospho-mTOR (S2448) Mouse Monoclonal Antibody [A5D5]

Antibody Type

Mouse Monoclonal Antibody

Immunogen

Synthetic peptide within human mTOR aa 2430-2480.

Species Reactivity

Human, Mouse (Predicted: Rat)

Predicted species support after-sales service

Validated Applications

WB, IHC-P

Molecular Weight

Predicted band size: 289 kDa

Positive Control

HeLa cell lysate, HeLa starved overnight then treated with 200nM PMA for 4 hours cell lysate, NIH/3T3 cell lysate treated with serum-starvation overnight, NIH/3T3 cell lysate treated with Insulin (100nM) for 10 minutes after serum-starvation overnight, human liver tissue.

Conjugation

unconjugated

Clone Number

A5D5

RRID

AB_3071710

同源性数据

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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

PBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG1

Purification Method

Protein G affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:2,000

  • IHC-P

  • 1:50-1:200

Target

Function

The phosphatidylinositol kinase (PIK) family members fall into two distinct subgroups. The first subgroup contains proteins such as the PI 3- and PI 4-kinases and the second group comprises the PIK-related kinases. The PIK-related kinases include Atm, DNA-PKCS and FRAP. These proteins have in common a region of homology at their carboxy-termini that is not present in the PI 3- and PI 4-kinases. The Atm gene is mutated in the autosomal recessive disorder ataxia telangiectasia (AT) that is characterized by cerebellar degeneration (ataxia) and the appearance of dilated blood vessels (telangiectases) in the conjunctivae of the eyes. AT cells are hypersensitive to ionizing radiation, impaired in mediating the inhibition of DNA synthesis and display delays in p53 induction. DNA-PK is a heterotrimeric DNA binding enzyme that is composed of a large subunit, DNA-PKCS, and two smaller subunits collectively known as Ku. The loss of DNA-PK leads to defects in DSB repair and V(D)J recombination. FRAP can autophosphorylate on serine and bind to Rapamycin/FKBP. FRAP is also an upstream regulator of S6 kinase and has been implicated in the regulation of p27 and p21 expression.

Background References

1. Kim D.-H. et. al. mTOR interacts with raptor to form a nutrient-sensitive complex that signals to the growth machinery. Cell 110:163-175(2002).

Subcellular Location

Lysosome, Mitochondrion outer membrane, Endoplasmic reticulum membrane, Cytoplasm.

UNIPROT

SwissProt: P42345 Human

SwissProt: Q9JLN9 Mouse

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

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-mTOR (S2448) on different lysates with Mouse anti-Phospho-mTOR (S2448) antibody (<a href="/products/HA600094" style="font-weight: bold;text-decoration: underline;">HA600094</a>) at 1/5,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: HeLa starved overnight then treated with 200nM PMA for 4 hours cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 289 kDa<br />Observed band size: 289 kDa<br /><br />Exposure time: 1 minute 30 seconds;<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/HA600094" style="font-weight: bold;text-decoration: underline;">HA600094</a>) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of Phospho-mTOR (S2448) on different lysates with Mouse anti-Phospho-mTOR (S2448) antibody (HA600094) at 1/5,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: HeLa starved overnight then treated with 200nM PMA for 4 hours cell lysate

    Lysates/proteins at 20 µg/Lane.

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

    Exposure time: 1 minute 30 seconds;

    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 (HA600094) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-mTOR (S2448) on different lysates with Mouse anti-Phospho-mTOR (S2448) antibody (<a href="/products/HA600094" style="font-weight: bold;text-decoration: underline;">HA600094</a>) at 1/1,000 dilution.<br /><br />Lane 1: NIH/3T3 cell lysate treated with serum-starvation overnight<br />Lane 2: NIH/3T3 cell lysate treated with Insulin (100nM) for 10 minutes after serum-starvation overnight<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 288 kDa<br />Observed band size: 255 kDa<br /><br />Exposure time: 2 minutes;<br /><br />10% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% BSA for 1 hour at room temperature. The primary antibody (<a href="/products/HA600094" style="font-weight: bold;text-decoration: underline;">HA600094</a>) at 1/1,000 dilution was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of Phospho-mTOR (S2448) on different lysates with Mouse anti-Phospho-mTOR (S2448) antibody (HA600094) at 1/1,000 dilution.

    Lane 1: NIH/3T3 cell lysate treated with serum-starvation overnight
    Lane 2: NIH/3T3 cell lysate treated with Insulin (100nM) for 10 minutes after serum-starvation overnight

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 288 kDa
    Observed band size: 255 kDa

    Exposure time: 2 minutes;

    10% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% BSA for 1 hour at room temperature. The primary antibody (HA600094) at 1/1,000 dilution was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human liver tissue with Mouse anti-Phospho-mTOR (S2448) antibody (<a href="/products/HA600094" style="font-weight: bold;text-decoration: underline;">HA600094</a>) at 1/100 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/HA600094" style="font-weight: bold;text-decoration: underline;">HA600094</a>) at 1/400 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 liver tissue with Mouse anti-Phospho-mTOR (S2448) antibody (HA600094) at 1/100 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 (HA600094) at 1/400 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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