Phospho-mTOR (Ser2448) Monoclonal antibody

Phospho-mTOR (Ser2448) Monoclonal Antibody for WB, IHC, IF/ICC, FC (Intra), ELISA

Host / Isotype

Mouse / IgG2b

Reactivity

human, mouse, rat and More (3)

Applications

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

Conjugate

Unconjugated

Publications

273

CloneNo.

2A12G3

Reviews (1)

Cat no : 67778-1-Ig

Synonyms

p mTOR, p mTOR (Ser2448), Phospho mTOR, Phospho mTOR (Ser2448), Phospho-mTOR

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Validation Data Gallery

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  WB analysis using 67778-1-Ig

  Non-treated and Calyculin A treated HeLa cells were subjected to SDS PAGE followed by western blot with 67778-1-Ig (Phospho-mTOR (Ser2448) antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 67778-1-Ig

  Non-treated and EGF treated NIH/3T3 cells were subjected to SDS PAGE followed by western blot with 67778-1-Ig (Phospho-mTOR (Ser2448) antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 67778-1-Ig

  Non-treated and Rapamycin treated HEK-293 cells were subjected to SDS PAGE followed by western blot with 67778-1-Ig (Phospho-mTOR (Ser2448) antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with RICTOR antibody (66867-2-Ig) and mTOR antibody (66888-1-Ig) subsequently.

• 

  WB analysis using 67778-1-Ig

  Non-treated and Calyculin A treated HEK-293 cells were subjected to SDS PAGE followed by western blot with 67778-1-Ig (Phospho-mTOR (Ser2448) antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with RICTOR antibody (66867-2-Ig) and mTOR antibody (66888-1-Ig) subsequently.

• 

  WB analysis using 67778-1-Ig

  Non-treated and Calyculin A treated HeLa cells were subjected to SDS PAGE followed by western blot with 67778-1-Ig (Phospho-mTOR (Ser2448) antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with RICTOR antibody (66867-2-Ig) and mTOR antibody (66888-1-Ig) subsequently.

• 

  WB analysis of HSC-T6 using 67778-1-Ig

  HSC-T6 cells were subjected to SDS PAGE followed by western blot with 67778-1-Ig (Phospho-mTOR (Ser2448) antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

• 

  IHC staining of human colon cancer using 67778-1-Ig

  Immunohistochemical analysis of paraffin-embedded human colon cancer tissue slide using 67778-1-Ig (Phospho-mTOR (Ser2448) antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human breast cancer using 67778-1-Ig

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 67778-1-Ig (Phospho-mTOR (Ser2448) antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human colon cancer using 67778-1-Ig

  Immunohistochemical analysis of paraffin-embedded human colon cancer tissue slide using 67778-1-Ig (Phospho-mTOR (Ser2448) antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human breast cancer using 67778-1-Ig

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 67778-1-Ig (Phospho-mTOR (Ser2448) antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IF Staining of HepG2 using 67778-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed HepG2 cells using Phospho-mTOR (Ser2448) antibody (67778-1-Ig, Clone: 2A12G3 ) at dilution of 1:200 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

• 

  FC experiment of HeLa using 67778-1-Ig

  1X10^6 Calyculin A treated HeLa cells were intracellularly stained with 0.5 ug Anti-Human Phospho-mTOR (Ser2448) (67778-1-Ig, Clone:2A12G3) and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (red), or 0.5 ug Control Antibody. Cells were fixed with 4% PFA and permeabilized with 90% MeOH.

• 

  FC experiment of HeLa using 67778-1-Ig

  1x10^6 100 nM Calyculin A (30 minutes) treated HeLa cells were intracellularly stained with 0.2 μg Phospho-mTOR (Ser2448) Monoclonal antibody (67778-1-Ig, Clone:2A12G3) and CoraLite® Plus 647-Goat Anti-Mouse Recombinant Secondary Antibody (H+L)(Cat.NO.RGAM005), and 0.2 μg KLH (66360-3-Ig, Clone: K11B8C4B5). Cells were fixed with 4% PFA .

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Tested Applications

• Positive WB detected in: HeLa cells, NIH/3T3 cells, HEK-293T cells, HEK-293 cells, HSC-T6 cells, Calyculin A treated HeLa cells, EGF treated NIH/3T3 cells, Rapamycin treated HEK-293 cells, Calyculin A treated HEK-293 cells
• Positive IHC detected in: human colon 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/ICC detected in: HepG2 cells
• Positive FC (Intra) detected in: Calyculin A treated HeLa cells, 100 nM Calyculin A (30 minutes) treated HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:2000-1:10000
• Immunohistochemistry (IHC): IHC : 1:500-1:2000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:50-1:500
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.50 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.

Published Applications

• WB: See 264 publications below
• IHC: See 17 publications below
• IF: See 14 publications below

Product Information

67778-1-Ig targets Phospho-mTOR (Ser2448) in WB, IHC, IF/ICC, FC (Intra), ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse, rat, pig, chicken, bovine
• Host / Isotype: Mouse / IgG2b
• Class: Monoclonal
• Type: Antibody
• Immunogen: Peptide
• Full Name: FK506 binding protein 12-rapamycin associated protein 1
• Calculated Molecular Weight: 289 kDa
• Observed Molecular Weight: 289 kDa
• GenBank Accession Number: BC117166
• Gene Symbol: mTOR
• Gene ID (NCBI): 2475
• RRID: AB_2889842
• 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.

Publications

Species	Application	Title
human	WB	Autophagy; Periplocin suppresses the growth of colorectal cancer cells by triggering LGALS3 (galectin 3)-mediated lysophagy; Authors - Kui Wang
mouse	WB	Redox Biol; GSTP1-mediated S-glutathionylation of Pik3r1 is a redox hub that inhibits osteoclastogenesis through regulating autophagic flux; Authors - Xiaoxiao Ji
human	WB	J Exp Clin Cancer Res; Circular RNA circATP9A promotes non-small cell lung cancer progression by interacting with HuR and by promoting extracellular vesicles-mediated macrophage M2 polarization; Authors - Yuanshan Yao
human	WB	J Control Release; cRGD-targeted heparin nanoparticles for effective dual drug treatment of cisplatin-resistant ovarian cancer; Authors - Xiaomei Liang
human	WB	Mater Today Bio; Carbon dots enhance extracellular matrix secretion for dentin-pulp complex regeneration through PI3K/Akt/mTOR pathway-mediated activation of autophagy.; Authors - Lili Liu
human	WB	J Adv Res; Salidroside sensitizes Triple-negative breast cancer to ferroptosis by SCD1-mediated lipogenesis and NCOA4-mediated ferritinophagy; Authors - Guiqin Huang

Reviews

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

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FH

Lilas (Verified Customer) (06-29-2021)

Used antibody at a 1:1000 dilution in 5% BSA, and overnight incubation at 4 degrees. A sharp band detected above the 245 kDA marker using western blot.

• Applications: Western Blot
• Primary Antibody Dilution: 1:1000
• Cell Tissue Type: Glioblastoma cell lines