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Phospho-mTOR (Ser2448) Monoclonal Antibody (MRRBY), eFluor™ 450, eBioscience™
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FIGURE: 1 / 5
Phospho-mTOR (Ser2448) Antibody (48-9718-41) in Flow
Normal human peripheral blood cells were unstimulated (left) or stimulated with Anti-Human CD3 and CD28 Functional Grade Purifieds (Product # 16-0037-81 and Product # 16-0289-81) for 48 hours (right). The cells were then intracellularly stained with Anti-Human CD3 APC (Product # 17-0036-42) and Anti-Human/Mouse phospho-mTOR (S2448) eFluor® 450 using the Intracellular Fixation & Permeabilization Buffer Set (... View More
Product Details
48-9718-41
Applications
Tested Dilution
Publications
Product Specifications
Published species
Mouse
Host/Isotype
Mouse
/ IgG2a, kappa
Recommended Isotype Control
Class
Monoclonal
Type
Antibody
Clone
MRRBY
Conjugate
Excitation/Emission Max
405/445 nm
View spectra
Form
Liquid
Concentration
5 µL/Test
Storage conditions
4°C, store in dark, DO NOT FREEZE!
Shipping conditions
Wet ice
RRID
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 because it 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 stimulated normal human peripheral blood cells. This can be used at 5 µL (0.06 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 BestProtocols 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 BestProtocols® Section under the Resources tab online.
eFluor™ 450 is an alternative to Pacific Blue™. eFluor™ 450 emits at 445 nm and is excited with the Violet laser (405 nm). Please make sure that your instrument is capable of detecting this fluorochrome.
Excitation: 405 nm; Emission: 445 nm; Laser: Violet 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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