Rabbit Anti-Phospho-mTOR (Ser2481) antibody

Home > Product > Antibody > Rabbit Anti-Phospho-mTOR (Ser2481) antibody

mTOR (phospho S2481); p-mTOR (phospho S2481); dJ576K7.1 (FK506 binding protein 12 rapamycin associated protein 1); FK506 binding protein 12 rapamycin associated protein 1; FK506 binding protein 12 rapamycin associated protein 2; FK506 binding protein 12 r

Cat:

SL3495R

Species Reactivity：

Human,Mouse,Rat,(predicted: Dog,Pig,Horse,Rabbit,Sheep,)

Immunogen：

KLH conjugated Synthesised phosphopeptide derived from human mTOR around the phosphorylation site of Ser2481:IH(p-S)FI

Format：

Liquid

Storage instructions：

Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.

Concentration：

1mg/ml

Clonality：

Polyclonal

Isotype：

IgG

Applications：

WB=1:500-2000ELISA=1:5000-10000IHC-P=1:100-500IHC-F=1:100-500Flow-Cyt=1μg /testICC=1:100IF=1:100-500（Paraffin sections need to do antigen repair）not yet tested in other applications.optimal dilutions/concentrations should be determined by the end user.

Host：

Rabbit

Product Overview：

Sample: Hepg2(Human) Cell Lysate at 30 ugPrimary: Anti-Phospho-mTOR (Ser2481) (SL3495R) at 1/300 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 289 kDObserved band size: 289 kDSample: Jurkat(Human) Cell Lysate at 30 ugMCF-7(Human) Cell Lysate at 30 ugPrimary: Anti- Phospho-mTOR (Ser2481) (SL3495R) at 1/1000 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 289 kDObserved band size: 289 kDParaformaldehyde-fixed, paraffin embedded (rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Phospho-mTOR (Ser2481) ) Polyclonal Antibody, Unconjugated (SL3495R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.Paraformaldehyde-fixed, paraffin embedded (mouse brain tissue); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Phospho-mTOR (Ser2481)) Polyclonal Antibody, Unconjugated (SL3495R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.Tissue/cell: rat brain tissue; 4% Paraformaldehyde-fixed and paraffin-embedded; Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,SLC0005) at 37℃ for 20 min; Incubation: Anti- Phospho-mTOR (Ser2481) Polyclonal Antibody, Unconjugated(SL3495R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) stainingA431 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, SLC0005) at 37°C for 20 min; Antibody incubation with (Phospho-mTOR (Ser2481)) polyclonal Antibody, Unconjugated (SL3495R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.Blank control:A431. Primary Antibody (green line): Rabbit Anti-Phospho-mTOR (Ser2481) antibody (SL3495R) Dilution: 1ug/Test; Secondary Antibody : Goat anti-rabbit IgG-AF488Dilution: 0.5ug/Test. ProtocolThe cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at -20℃.The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.Blank control (blue line): Hela(fixed with 70% ethanol (Overnight at 4℃) and then permeabilized with 90% ice-cold methanol for 30 min on ice) Primary Antibody (green line): Rabbit Anti-Phospho-mTOR (Ser2481) antibody (SL3495R),Dilution: 1μg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG . Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC, Dilution: 1μg /test. Blank control: mouse splenocytes(blue)Isotype Control Antibody: Rabbit IgG(orange) ; Secondary Antibody: Goat anti-rabbit IgG-FITC(white blue), Dilution: 1:100 in 1 X PBS containing 0.5% BSA ; Primary Antibody Dilution: 5μl in 100 μL1X PBS containing 0.5% BSA(green).

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Unit:

Price: $256.00

Product PDFs

Datasheet: Down Datasheet

Other Information
Citations
Protein Attributes
Feedback Wall

mTOR is one of a family of proteins involved in cell cycle progression, DNA recombination, and DNA damage detection. In rat, it is a 289-kDa protein (symbolized RAFT1) with significant homology to the Saccharomyces cerevisiae protein TOR1 and has been shown to associate with the immunophilin FKBP12 in a rapamycin dependent fashion. The FKBP12-rapamycin complex is known to inhibit progression through the G1 cell cycle stage by interfering with mitogenic signaling pathways involved in G1 progression in several cell types, as well as in yeast. The binding of FRAP to FKBP12-rapamycin correlated with the ability of these ligands to inhibit cell cycle progression.

Function:
Serine/threonine protein kinase which is a central regulator of cellular metabolism, growth and survival in response to hormones, growth factors, nutrients, energy and stress signals. Functions as part of 2 structurally and functionally distinct signaling complexes mTORC1 and mTORC2 (mTOR complex 1 and 2). Activated mTORC1 up-regulates protein synthesis by phosphorylating key regulators of mRNA translation and ribosome synthesis. This includes phosphorylation of EIF4EBP1 and release of its inhibition toward the elongation initiation factor 4E (eiF4E). Moreover, phosphorylates and activates RPS6KB1 and RPS6KB2 that promote protein synthesis by modulating the activity of their downstream targets including ribosomal protein S6, eukaryotic translation initiation factor EIF4B and the inhibitor of translation initiation PDCD4. Regulates ribosome synthesis by activating RNA polymerase III-dependent transcription through phosphorylation and inhibition of MAF1 a RNA polymerase III-repressor. In parallel to protein synthesis, also regulates lipid synthesis through SRESF1/SREBP1 and LPIN1. To maintain energy homeostasis mTORC1 may also regulate mitochondrial biogenesis through regulation of PPARGC1A. mTORC1 also negatively regulates autophagy through phosphorylation of ULK1. Under nutrient sufficiency, phosphorylates ULK1 at 'Ser-758', disrupting the interaction with AMPK and preventing activation of ULK1. Also prevents autophagy through phosphorylation of the autophagy inhibitor DAP. mTORC1 exerts a feedback control on upstream growth factor signaling that includes phosphorylation and activation of GRB10 a INSR-dependent signaling suppressor. Among other potential targets mTORC1 may phosphorylate CLIP1 and regulate microtubules. As part of the mTORC2 complex MTOR may regulate other cellular processes including survival and organization of the cytoskeleton. Plays a critical role in the phosphorylation at 'Ser-473' of AKT1, a pro-survival effector of phosphoinositide 3-kinase, facilitating its activation by PDK1. mTORC2 may regulate the actin cytoskeleton, through phosphorylation of PRKCA, PXN and activation of the Rho-type guanine nucleotide exchange factors RHOA and RAC1A or RAC1B. mTORC2 also regulates the phosphorylation of SGK1 at 'Ser-422'.

Subunit:
Part of the mammalian target of rapamycin complex 1 (mTORC1) which contains MTOR, MLST8, RPTOR, AKT1S1/PRAS40 and DEPTOR. Part of the mammalian target of rapamycin complex 2 (mTORC2) which contains MTOR, MLST8, PRR5, RICTOR, MAPKAP1 and DEPTOR. Interacts with PPAPDC3 and PML (By similarity). Interacts with PRR5 and RICTOR; the interaction is direct within the mTORC2 complex (By similarity). Interacts with UBQLN1 (By similarity). Interacts with TTI1 and TELO2 (By similarity). Interacts with CLIP1; phosphorylates and regulates CLIP1 (By similarity).

Subcellular Location:
Endoplasmic reticulum membrane; Peripheral membrane protein; Cytoplasmic side. Golgi apparatus membrane; Peripheral membrane protein; Cytoplasmic side. Mitochondrion outer membrane; Peripheral membrane protein; Cytoplasmic side. Lysosome. Cytoplasm (By similarity). Nucleus, PML body (By similarity). Note=Shuttles between cytoplasm and nucleus. Accumulates in the nucleus in response to hypoxia (By similarity). Targeting to lysosomes depends on amino acid availability and RRAGA and RRAGB.

Tissue Specificity:
Expressed in numerous tissues, with highest levels in testis.

Post-translational modifications:
Phosphorylated. Autophosphorylates when part of mTORC1 or mTORC2. Phosphorylation at Ser-1261 promotes autophosphorylation.

Similarity:
Belongs to the PI3/PI4-kinase family.
Contains 1 FAT domain.
Contains 1 FATC domain.
Contains 7 HEAT repeats.
Contains 1 PI3K/PI4K domain.

SWISS:
P42345

Gene ID:
2475

Database links:

Entrez Gene: 2475 Human

Entrez Gene: 56717 Mouse

Entrez Gene: 56718 Rat

Omim: 601231 Human

SwissProt: P42345 Human

SwissProt: Q9JLN9 Mouse

SwissProt: P42346 Rat

Unigene: 338207 Human

Unigene: 21158 Mouse

Unigene: 11008 Rat

mTOR蛋白是一种丝氨酸／苏氨酸蛋白激酶（Ser/Thr），是调节细胞生长和增殖的重要信号转导分子，在细胞的生长、分化、增殖、迁移和存活上扮演了重要的角色。由于mTOR信号通路在细胞周期进程中发挥了重要作用，细胞周期进程调节异常与许多疾病尤其是癌症的发生、发展有关，mTOR信号通路的失调可引起多种肿瘤的发生。 Picture

Sample:
Jurkat(Human) Cell Lysate at 30 ug
MCF-7(Human) Cell Lysate at 30 ug
Primary: Anti- Phospho-mTOR (Ser2481) (SL3495R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 289 kD
Observed band size: 289 kD

Paraformaldehyde-fixed, paraffin embedded (rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Phospho-mTOR (Ser2481) ) Polyclonal Antibody, Unconjugated (SL3495R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (mouse brain tissue); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Phospho-mTOR (Ser2481)) Polyclonal Antibody, Unconjugated (SL3495R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Tissue/cell: rat brain tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,SLC0005) at 37℃ for 20 min;
Incubation: Anti- Phospho-mTOR (Ser2481) Polyclonal Antibody, Unconjugated(SL3495R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) staining

A431 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, SLC0005) at 37°C for 20 min; Antibody incubation with (Phospho-mTOR (Ser2481)) polyclonal Antibody, Unconjugated (SL3495R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.

Blank control:A431.
Primary Antibody (green line): Rabbit Anti-Phospho-mTOR (Ser2481) antibody (SL3495R)
Dilution: 1ug/Test;
Secondary Antibody : Goat anti-rabbit IgG-AF488
Dilution: 0.5ug/Test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at -20℃.The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.

Blank control (blue line): Hela(fixed with 70% ethanol (Overnight at 4℃) and then permeabilized with 90% ice-cold methanol for 30 min on ice)
Primary Antibody (green line): Rabbit Anti-Phospho-mTOR (Ser2481) antibody (SL3495R),Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC, Dilution: 1μg /test.

Blank control: mouse splenocytes(blue)
Isotype Control Antibody: Rabbit IgG(orange) ; Secondary Antibody: Goat anti-rabbit IgG-FITC(white blue), Dilution: 1:100 in 1 X PBS containing 0.5% BSA ; Primary Antibody Dilution: 5μl in 100 μL1X PBS containing 0.5% BSA(green).

Product Feedback Wall

Specific References (8) | SL3495R has been referenced in 8 publications.

[IF=3.848] Chen Luyao. et al. Epicatechin gallate prevents the de novo synthesis of fatty acid and the migration of prostate cancer cells. Acta Bioch Bioph Sin. 2021 Oct;: WB ; Human.

PubMed:34718375

[IF=4.451] Lei Wang. et al. Neuroprotective effect of Lactobacillus plantarum DP189 on MPTP-induced Parkinson's disease model mice. J Funct Foods. 2021 Oct;85:104635 WB ; Mouse.

PubMed:10.1016/j.jff.2021.104635

[IF=3.647] Shuang Huet al. MicroRNA-708 prevents ethanol-induced hepatic lipid accumulation and inflammatory reaction via direct targeting ZEB1. Life Sci . 2020 Oct 1;258:118147. WB ; Human.

PubMed:32721464

[IF=3.448] Jin Y et al. miR‐496 remedies hypoxia reoxygenation–induced H9c2 cardiomyocyte apoptosis via Hook3‐targeted PI3k/Akt/mTOR signaling pathway activation. J Cell Biochem. 2019 Aug 22. WB ; Rat&Human.

PubMed:31436348

[IF=3.04] Shi C et al. Acetaminophen aggravates fat accumulation in NAFLD by inhibiting autophagy via the AMPK/mTOR pathway. European Journal of Pharmacology.2019. WB&IHSLCP ; Mouse&Human.

PubMed:10.1016/j.ejphar.2019.02.005

[IF=2.976] Guo H et al. Clinical associations between ASCT2 and p-mTOR in the pathogenesis and prognosis of epithelial ovarian cancer. Oncol Rep. 2018 Dec;40(6):3725-3733. WB&IHSLCP ; Human.

PubMed:30272366

[IF=4.527] Zhao HJ et al. Destruction of redox and mitochondrial dynamics co-contributes to programmed cell death in chicken kidney under arsenite or/and copper (II) exposure. Ecotoxicology and Environmental Safety, 2019 179, 167–174. WB ; Chicken.

PubMed:doi:10.1016/j.ecoenv.2019.04.062

[IF=3.414] Ma HX et al. Mu-Xiang-You-Fang protects PC12 cells against OGD/R-induced autophagy via the AMPK/mTOR signaling pathway. J Ethnopharmacol. 2020 Jan 22;252:112583. WB ; Rattus norvegicus.

PubMed:31978519