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

货号：46-9718-41,46-9718-42

规格：25 Tests,100 Tests

价格：0,3783

产品类型：流式抗体

品牌：Thermo Fisher

物种：人/小鼠

宿主：小鼠

抗体亚型：IgG2a, kappa

克隆号：MRRBY

荧光染料：PerCP-eFluor™ 710

类型:	单抗	同型对照：	Mouse IgG2a kappa Isotype Control (eBM2a), PerCP-eFluor 710
浓度：	5 µL/Test	用法：	5 µL (0.125 µg)/test(Flow)

产品详细信息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 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 (cat. 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 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.靶标信息Mammalian target of rapamycin (mTOR) is a serine/threonine kinase that plays a key role in cell growth, cell proliferation, and protein synthesis. mTOR mediates phosphoinositide 3-kinase and Akt/PKB signaling, resulting in phosphorylation of 4EBP1, and initiation of mRNA translation. A second pathway involves regulation of ribosomal S6 kinase, which affects ribosome biogenesis and translation elongation. Mutations affecting the gene results in Smith-Kingsmore syndrome.

数据

Phospho-mTOR (Ser2448) Antibody (46-9718-42) in FlowTOP: Normal human peripheral blood cells were unstimulated (left) or stimulated with Anti-Human CD3 Functional Grade Purified (Product # 16-0037-81) plus Anti-Human CD28 Functional Grade Purified (Product # 16-0289-81) (right) for 48 hours. The cells were then intracellularly stained with Anti-Human CD3 FITC (Product # 11-0036-42) and Anti-Human/Mouse phospho-mTOR (S2448) PerCP-eFluor 710 using the Intracellular Fixation and Permeabilization Buffer Set (Product # 88-8824-00) and protocol. Cells in the lymphocyte gate were used for analysis. BOTTOM: Normal human peripheral blood cells were unstimulated (orange histogram) or were stimulated with Anti-Human CD3 Functional Grade Purified (Product # 16-0037-81) plus Anti-Human CD28 Functional Grade Purified (Product # 16-0289-81) in the presence (green histogram) or absence (purple histogram) of the PI3 kinase inhibitor, LY294002, for 48 hours. The cells were then intracellularly stained with Anti-Human CD3 FITC (Product # 11-0036-42) and Anti-Human/Mouse phospho-mTOR (S2448) PerCP-eFluor 710 using the Intracellular Fixation and Permeabilization Buffer Set (Product # 88-8824-00) and protocol. CD3+ cells in the lymphocyte gate were used for analysis.

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