Invitrogen

Phospho-mTOR (Ser2448) Monoclonal Antibody (MRRBY), PerCP-eFluor™ 710, eBioscience™

3 Published Figures
3 References
View all (96) mTOR antibodies
Datasheet
Protocols
Questions & Answers
Datasheet
Protocols
Questions & Answers

Product Details

46-9718-42

Applications
Tested Dilution
Publications

Flow Cytometry (Flow)

5 µL (0.125 µg)/test
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Product Specifications

Species Reactivity

Human, Mouse

Published species

Mouse

Host/Isotype

Mouse / IgG2a, kappa

Recommended Isotype Control

Mouse IgG2a kappa Isotype Control (eBM2a), PerCP-eFluor™ 710, eBioscience™

Class

Monoclonal

Type

Antibody

Clone

MRRBY

Conjugate

PerCP-eFluor™ 710 PerCP-eFluor™ 710 PerCP-eFluor™ 710

Excitation/Emission Max

482/708 nm View spectra spectra

Form

Liquid

Concentration

5 µL/Test

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2, with 0.2% BSA

Contains

0.09% sodium azide

Storage conditions

4° C, store in dark, DO NOT FREEZE!

Shipping conditions

Ambient (domestic); Wet ice (international)

RRID

AB_2573895

Product Specific Information

Description: This MRRBY monoclonal antibody recognizes human and mouse mammalian target of rapamycin (also known as mTOR, FRAP, RAFT) when phosphorylated on S2448. mTOR is a serine/threonine protein kinase that functions as an ATP and amino acid sensor as well as to balance nutrient availability with cell growth, proliferation, motility, survival, protein synthesis, and transcription. Activated mTOR increases production of enzymes necessary for glycolysis and controls the uptake of glucose and other nutrients. Increased glucose uptake and metabolism helps fulfill the energy needs for mTOR-driven cell growth and proliferation. When sufficient nutrients are available, mTOR transmits a positive signal to p70 S6 kinase and participates in the inactivation of the eIF4E inhibitor, 4E-BP1. mTOR is phosphorylated at S2448 via the PI3 kinase/Akt signaling pathway and is autophosphorylated at Ser2481. Due to its critical role in regulation of cell growth, survival, and metabolism, and becauseit is often abnormally regulated in tumors, mTOR is under investigation as a potential target for anti-cancer therapy.

Applications Reported: This MRRBY antibody has been reported for use in intracellular staining followed by flow cytometric analysis.

Applications Tested: This MRRBY antibody has been pre-titrated and tested by intracellular staining followed by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (0.125 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.

Staining Protocol: All protocols work well for this monoclonal antibody. Use of Protocol A: Two-step protocol: intracellular (cytoplasmic) proteins allows for the greatest flexibility for detection of surface and intracellular (cytoplasmic) proteins. Use of Protocol B: One-step protocol: intracellular (nuclear) proteins is recommended for staining of transcription factors in conjunction with surface and phosphorylated intracellular (cytoplasmic) proteins. Protocol C: Two-step protocol: Fixation/Methanol allows for the greatest discrimination of phospho-specific signaling between unstimulated and stimulated samples, but with limitations on the ability to stain specific surface proteins (refer to "Clone Performance Following Fixation/Permeabilization" located in the Best Protocols Section under the Resources tab online). All Protocols can be found in the Flow Cytometry Protocols: "Staining Intracellular Antigens for Flow Cytometry Protocol" located in the Best Protocols Section under the Resources tab online.

PerCP-eFluor® 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome.

Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.

Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.

Excitation: 488 nm; Emission: 710 nm; Laser: Blue Laser.

Filtration: 0.2 µm post-manufacturing filtered.

Target Information

FRAP1 (mTOR) is a serine/threonine kinase that plays a critical role in cellular growth and proliferation. Perturbations in the mTOR/PI3-kinase/AKT pathway are associated with numerous forms of cancer. FRAP1 is also the target of rapamycin and its analogues, which are currently used as immunosuppressants and cancer therapeutics. Mutations affecting the gene results in Smith-Kingsmore syndrome.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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Bioinformatics

Protein Aliases: angiopoietin-like factor CDT6; FK506 binding protein 12-rapamycin associated protein 1; FK506 binding protein 12-rapamycin associated protein 2; FK506-binding protein 12-rapamycin complex-associated protein 1; FKBP-rapamycin associated protein; FKBP-rapamycin associated protein (FRAP); FKBP-rapamycin-associated protein FRAP; FKBP12-rapamycin complex-associated protein; FKBP12-rapamycin complex-associated protein 1; m-TOR; Mammalian target of rapamycin; Mechanistic target of rapamycin; mechanistic target of rapamycin (serine/threonine kinase); mTOR; mTORC1; Rapamycin and FKBP12 target 1; rapamycin associated protein FRAP2; Rapamycin target protein 1; RAPT1; Serine/threonine-protein kinase mTOR

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Gene Aliases: 2610315D21Rik; AI327068; flat; FRAP; FRAP1; FRAP2; MTOR; RAFT1; RAPT1; SKS

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UniProt ID: (Human) P42345, (Mouse) Q9JLN9

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Entrez Gene ID: (Human) 2475, (Mouse) 56717

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Function(s)
RNA polymerase III type 1 promoter DNA binding RNA polymerase III type 2 promoter DNA binding RNA polymerase III type 3 promoter DNA binding TFIIIC-class transcription factor binding protein kinase activity protein serine/threonine kinase activity protein binding ATP binding kinase activity protein kinase binding protein domain specific binding ribosome binding phosphoprotein binding nucleotide binding drug binding transferase activity transferase activity, transferring phosphorus-containing groups phosphotransferase activity, alcohol group as acceptor protein dimerization activity non-receptor serine/threonine protein kinase protein modifying enzyme
Process(es)
negative regulation of protein phosphorylation positive regulation of protein phosphorylation positive regulation of endothelial cell proliferation heart morphogenesis heart valve morphogenesis regulation of glycogen biosynthetic process energy reserve metabolic process 'de novo' pyrimidine nucleobase biosynthetic process DNA repair protein phosphorylation response to stress cell cycle arrest signal transduction germ cell development brain development cell aging response to nutrient long-term memory visual learning post-embryonic development negative regulation of autophagy positive regulation of lamellipodium assembly positive regulation of gene expression positive regulation of myotube differentiation positive regulation of neuron maturation negative regulation of muscle atrophy cell growth macroautophagy negative regulation of macroautophagy phosphorylation peptidyl-serine phosphorylation peptidyl-threonine phosphorylation spinal cord development protein catabolic process positive regulation of actin filament polymerization T cell costimulation negative regulation of protein ubiquitination ruffle organization regulation of myelination cellular response to nutrient levels TOR signaling regulation of fatty acid beta-oxidation regulation of response to food response to insulin regulation of actin cytoskeleton organization social behavior multicellular organism growth growth wound healing response to cocaine regulation of GTPase activity response to amino acid response to morphine regulation of carbohydrate utilization positive regulation of nitric oxide biosynthetic process regulation of osteoclast differentiation positive regulation of translation negative regulation of cell size regulation of protein kinase activity positive regulation of transcription from RNA polymerase III promoter protein autophosphorylation positive regulation of lipid biosynthetic process phosphatidylinositol-mediated signaling mRNA stabilization positive regulation of smooth muscle cell proliferation positive regulation of oligodendrocyte differentiation positive regulation of peptidyl-tyrosine phosphorylation voluntary musculoskeletal movement positive regulation of stress fiber assembly negative regulation of NFAT protein import into nucleus positive regulation of protein kinase B signaling cardiac muscle cell development cardiac muscle contraction maternal process involved in female pregnancy positive regulation of glial cell proliferation positive regulation of dendritic spine development positive regulation of cell growth involved in cardiac muscle cell development cellular response to hypoxia regulation of brown fat cell differentiation regulation of membrane permeability regulation of cellular response to heat positive regulation of neuron death positive regulation of transcription of nuclear large rRNA transcript from RNA polymerase I promoter positive regulation of eating behavior positive regulation of cholangiocyte proliferation positive regulation of sensory perception of pain negative regulation of cholangiocyte apoptotic process positive regulation of granulosa cell proliferation positive regulation of skeletal muscle hypertrophy negative regulation of iodide transmembrane transport double-strand break repair via homologous recombination regulation of carbohydrate metabolic process cell projection organization
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