mTOR Polyclonal antibody

• Featured Product
• KD/KO Validated

mTOR Polyclonal Antibody for WB, IP, IF, IHC, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

Human and More (6)

Applications

WB, IP, IF, IHC, ELISA

Conjugate

Unconjugated

Publications

78

Reviews (1)

Cat no : 28273-1-AP

Synonyms

FLJ44809, FRAP, FRAP1, FRAP2, Mammalian target of rapamycin, MTOR, RAFT1, Rapamycin target protein 1, RAPT1

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Validation Data Gallery

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  WB analysis of HeLa using 28273-1-AP

  WB result of mTOR antibody (28273-1-AP; 1:4000; incubated at room temperature for 1.5 hours) with sh-Control and sh-mTOR transfected HeLa cells.

• 

  WB analysis using 28273-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 28273-1-AP (mTOR antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 28273-1-AP

  Non-treated and Rapamycin treated lysates were subjected to SDS PAGE followed by western blot with 28881-1-AP (Phospho-mTOR (Ser2448) antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with mTOR antibody (28273-1-AP) and Vinculin (66305-1-Ig) subsequently.

• 

  WB analysis using 28273-1-AP

  HepG2 cells were subjected to SDS PAGE followed by western blot with 28273-1-AP (MTOR antibody) at dilution of 1:2500 incubated at room temperature for 1.5 hours.

• 

  WB analysis of PC-3 using 28273-1-AP

  PC-3 cells were subjected to SDS PAGE followed by western blot with 28273-1-AP (MTOR antibody) at dilution of 1:1500 incubated at room temperature for 1.5 hours.

• 

  WB analysis of MCF-7 using 28273-1-AP

  MCF-7 cells were subjected to SDS PAGE followed by western blot with 28273-1-AP (MTOR antibody) at dilution of 1:1500 incubated at room temperature for 1.5 hours.

• 

  WB analysis of HepG2 using 28273-1-AP

  HepG2 cells were subjected to SDS PAGE followed by western blot with 28273-1-AP (MTOR antibody) at dilution of 1:1500 incubated at room temperature for 1.5 hours.

• 

  IP experiment of HeLa using 28273-1-AP

  IP result of anti-mTOR (IP:28273-1-AP, 4ug; Detection:28273-1-AP 1:5000) with HeLa cells lysate 1720 ug.

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  IHC staining of human prostate cancer using 28273-1-AP

  Immunohistochemical analysis of paraffin-embedded human prostate cancer tissue slide using 28273-1-AP (MTOR antibody) at dilution of 1:200 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human prostate cancer using 28273-1-AP

  Immunohistochemical analysis of paraffin-embedded human prostate cancer tissue slide using 28273-1-AP (MTOR antibody) at dilution of 1:200 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human breast cancer using 28273-1-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 28273-1-AP (MTOR antibody) at dilution of 1:200 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human breast cancer using 28273-1-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 28273-1-AP (MTOR antibody) at dilution of 1:200 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IF Staining of HeLa using 28273-1-AP

  Immunofluorescent analysis of (-20°C Ethanol) fixed HeLa cells using mTOR antibody (28273-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L), CL594-Phalloidin (red).

• 

  IF Staining of HepG2 using 28273-1-AP

  Immunofluorescent analysis of (-20°C Ethanol) fixed HepG2 cells using mTOR antibody (28273-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L), CL594-Phalloidin (red).

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Tested Applications

• Positive WB detected in: HeLa cells, HepG2 cells, Rapamycin treated HEK-293T cells, PC-3 cells, MCF-7 cells, Rapamycin treated MCF-7 cells
• Positive IP detected in: HeLa cells
• Positive IHC detected in: human prostate cancer tissue, human breast cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF detected in: HeLa cells, HepG2 cells

Recommended dilution

• Western Blot (WB): WB : 1:2000-1:10000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:50-1:500
• Immunofluorescence (IF): IF : 1:50-1:500

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• KD/KO: See 1 publications below
• WB: See 78 publications below
• IHC: See 3 publications below
• IF: See 1 publications below

Product Information

28273-1-AP targets mTOR in WB, IP, IF, IHC, ELISA applications and shows reactivity with Human samples.

• Tested Reactivity: Human
• Cited Reactivity: human, mouse, rat, chicken, monkey, bovine
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: mTOR fusion protein Ag28395
• Full Name: FK506 binding protein 12-rapamycin associated protein 1
• Calculated Molecular Weight: 289 kDa
• Observed Molecular Weight: 250-289 kDa
• GenBank Accession Number: NM_004958
• Gene Symbol: FRAP1
• Gene ID (NCBI): 2475
• RRID: AB_2881102
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

MTOR, also named as FRAP1, FRAP, FRAP2 and RAPT1, belongs to the PI3/PI4-kinase family. MTOR is a Ser/Thr protein kinase that functions as an ATP and amino acid sensor to balance nutrient availability and cell growth. MTOR is kinase subunit of both mTORC1 and mTORC2, which regulate cell growth and survival in response to nutrient and hormonal signals. mTORC1 is activated in response to growth factors or amino-acids. mTORC2 is also activated by growth factors, but seems to be nutrient-insensitive. mTORC2 seems to function upstream of Rho GTPases to regulate the actin cytoskeleton, probably by activating one or more Rho-type guanine nucleotide exchange factors. mTORC2 promotes the serum-induced formation of stress-fibers or F-actin. MTOR has a calculated molecular mass of 289 kDa, and always can be detected at about 250 kDa due to some modifications (PMID: 14578359).

Publications

Species	Application	Title
mouse	WB	Proc Natl Acad Sci U S A; Heterozygous LRP1 deficiency causes developmental dysplasia of the hip by impairing triradiate chondrocytes differentiation due to inhibition of autophagy; Authors - Wenjin Yan
human	WB	J Transl Med; Fasting mimicking diet inhibits tumor-associated macrophage survival and pro-tumor function in hypoxia: implications for combination therapy with anti-angiogenic agent; Authors - Lei Wang
human	WB	Inflamm Res; Mammalian STE20-like kinase 1 inhibits synoviocytes activation in rheumatoid arthritis through mitochondrial dysfunction mediated by SIRT3/mTOR axis; Authors - Min Tan
mouse	WB	J Nanobiotechnology; Regulating immune memory and reversing tumor thermotolerance through a step-by-step starving-photothermal therapy; Authors - Lihua Luo
human	WB	Int J Mol Sci; Piperine Improves Lipid Dysregulation by Modulating Circadian Genes Bmal1 and Clock in HepG2 Cells.; Authors - Weiyun Zhang
mouse	WB	Free Radic Biol Med; Advanced oxidation protein products impair autophagic flux in macrophage by inducing lysosomal dysfunction via activation of PI3K-Akt-mTOR pathway in Crohn's disease.; Authors - Yan Liao

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive forproviding their feedback.

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FH

Tianyi (Verified Customer) (01-12-2024)

Antigen retrieval in TE buffer, incubated with AF594 at RT for 2h.

• Applications: Immunofluorescence
• Primary Antibody Dilution: 1:200
• Cell Tissue Type: Ovarian tissue