Phospho-AKT1 (Ser473) Monoclonal Antibody (SDRNR), PE

货号：12-9715-41,12-9715-42

规格：25 Tests,100 Tests

价格：1237,3481

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：SDRNR

荧光染料：PE

类型:	单抗	同型对照：	Mouse IgG2a kappa Isotype Control (eBM2a), PE
浓度：	5 µL/Test	用法：	5 µL (0.06 µg)/test(Flow)

产品详细信息Description: This SDRNR monoclonal antibody recognizes human and mouse AKT (also known as Protein Kinase B (PKB)) when phosphorylated on S473. AKT is a serine/threonine protein kinase that plays a key role in multiple cellular processes including metabolism, proliferation, apoptosis/survival, and migration. There are three homologous isoforms of AKT: AKT1, AKT2, and AKT3. AKT is activated by binding of its pleckstrin homology (PH) domain to membrane phospholipids and by phosphorylation. Phosphorylation of AKT at T308 by PDK1 and at S473 is required for full activation of this kinase. AKT promotes cell survival by inhibiting apoptosis via phosphorylation and inactivation of several targets including Bad, Foxo1, c-Raf, and caspase-9. Deregulation of AKT has been implicated as a major contributing factor in many types of cancer. AKT is negatively regulated by the phosphatase PTEN as well as by the chemical inhibitor LY294002. Specificity of this SDRNR clone was determined by ELISA, flow cytometry, and western blotting.Applications Reported: This SDRNR antibody has been reported for use in intracellular staining followed by flow cytometric analysis.Applications Tested: This SDRNR 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 (0.06 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 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.Excitation: 488-561 nm; Emission: 578 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser.Filtration: 0.2 µm post-manufacturing filtered.靶标信息AKT also known as protein kinase B (PKB) or RAS-alpha, is an ubiquitous serine/threonine kinase that plays an important role in diverse biological responses such as regulation of metabolism, cell survival and growth by phosphorylating multiple proteins. This protein kinase is activated by insulin, PI3K, IGF1 and various other growth and survival factors. Akt promotes cell survival by inhibiting apoptosis through phosphorylation and inactivation of several targets, including forkhead transcription factors, and caspase-9. The AKT pathway is a major target for cancer drug discovery.

数据

Phospho-AKT1 (Ser473) Antibody (12-9715-42)Intracellular staining of stimulated human peripheral blood cells. As expected based on known expression patterns, phospho-AKT (S473) clone SDRNR stains monocytes in response to LPS (right) treatment but does not stain lymphocytes (middle). Details: Normal human peripheral blood cells were unstimulated (purple histogram) or stimulated with Lipopolysaccharide (LPS) Solution (500X) (orange histogram), then intracellularly stained with Anti-Human/Mouse phospho-AKT (S473) (clone SDRNR) using the Intracellular Fixation and Permeabilization Buffer Set and protocol. Single cells in the lymphocyte (middle) and monocyte (right) gates were used for analysis. Staining of the isotype control, Mouse IgG2a kappa, on LPS-stimulated cells is shown in both plots (blue histogram).

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参考文献：

1.Nature communicationsCrosstalks between mTORC1 and mTORC2 variagate cytokine signaling to control NK maturation and effector function."Published figure using Phospho-AKT1 (Ser473) monoclonal antibody (Product # 12-9715-42) in Flow Cytometry"

技术参数

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