mTOR Recombinant antibody

• Featured Product
• KD/KO Validated

mTOR Uni-rAb^TM Recombinant Antibody for WB, IHC, IF/ICC, FC (Intra), IP, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat

Applications

WB, IHC, IF/ICC, FC (Intra), IP, ELISA

Conjugate

Unconjugated

CloneNo.

6H23

Cat no : 81670-1-RR

Synonyms

FRAP1, FRAP2, FRAP, FKBP12-rapamycin complex-associated protein, EC:2.7.11.1

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Validation Data Gallery

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  WB analysis of HeLa using 81670-1-RR

  WB result of mTOR antibody (81670-1-RR; 1:3000; incubated at room temperature for 1.5 hours) with sh-Control and sh-mTOR transfected HeLa cells.

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  WB analysis using 81670-1-RR

  Various lysates were subjected to SDS PAGE followed by western blot with 81670-1-RR (mTOR antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

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  IP experiment of HeLa using 81670-1-RR

  IP result of anti-mTOR (IP:81670-1-RR, 4ug; Detection:81670-1-RR 1:1000) with HeLa cells lysate 1760 ug.

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  IHC staining of human colon cancer using 81670-1-RR

  Immunohistochemical analysis of paraffin-embedded human colon cancer tissue slide using 81670-1-RR (mTOR antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of HepG2 using 81670-1-RR

  Immunofluorescent analysis of (-20°C Methanol) fixed HepG2 cells using mTOR antibody (81670-1-RR, Clone: 6H23 ) at dilution of 1:200 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L).

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  IF Staining of HepG2 using 81670-1-RR

  Immunofluorescent analysis of (-20°C Ethanol) fixed HepG2 cells using mTOR antibody (81670-1-RR, Clone: 6H23 ) at dilution of 1:200 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L), CL594-Phalloidin (red).

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  FC experiment of HeLa using 81670-1-RR

  1X10^6 HeLa cells were intracellularly stained with 0.4 ug Anti-Human mTOR (81670-1-RR, Clone:6H23) and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Isotype Control. Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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  Affinity and Kinetic Characterization of 81670-1-RR

  Biolayer interferometry (BLl) kinetic assays of 81670-1-RR against Human mTOR were performed. The affinity constant is below 1 pM.

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Tested Applications

• Positive WB detected in: HeLa cells, HEK-293 cells, MCF-7 cells, HepG2 cells, Jurkat cells, NIH/3T3 cells, HSC-T6 cells
• Positive IP detected in: HeLa cells
• Positive IHC detected in: human colon 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/ICC detected in: HepG2 cells
• Positive FC (Intra) detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:1000-1:6000
• 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)/ICC: IF/ICC : 1:50-1:500
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

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.

Product Information

81670-1-RR targets mTOR in WB, IHC, IF/ICC, FC (Intra), IP, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• 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: mTOR
• Gene ID (NCBI): 2475
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A 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 is phosphorylated at Ser2448 via the PI3 kinase/Akt signaling pathway and autophosphorylated at Ser2481. mTOR plays a key role in cell growth and homeostasis and may be abnormally regulated in tumors.