- 询价
- Cell Signaling Technology
- 4058
- USA
- 2025年10月03日
- western blot,免疫沉淀（IP）,免疫荧光（IF）,流式细胞（Flow Cyt）
- 兔
- 人,小鼠,大鼠
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- 详细信息
- 文献和实验
- 技术资料
- 抗体英文名：
Phospho-Akt (Ser473) (193H12) Rabbit mAb
- 抗原：
/
- 应用范围：
western blot,免疫沉淀（IP）,免疫荧光（IF）,流式细胞（Flow Cyt）
- 宿主：
兔
- 抗原来源：
/
- 保质期：
详见说明书
- 库存：
大量
- 适应物种：
人,小鼠,大鼠
- 级别：
详见MSDS文件
- 供应商：
CST
- 是否单克隆：
1
- 保存条件：
-20°c
- 规格：
100 ul (10 western blots)/300 ul (30 western blots)/<a href="http://www.cellsignal.com/ddt/custom_reagents.html" target="_blank">carrier free & custom formulation / quantity</a>
| 规格： | 产品价格： | ￥请询价 | |
|---|---|---|---|
| 规格： | 100 ul (10 western blots) | 产品价格： | ￥请询价 |
| 规格： | 300 ul (30 western blots) | 产品价格： | ￥请询价 |
| 规格： | <a href="http://www.cellsignal.com/ddt/custom_reagents.html" target="_blank">carrier free & custom formulation / quantity</a> | 产品价格： | ￥请询价 |
Product Pathways - PI3K / Akt Signaling
Phospho-Akt (Ser473) (193H12) Rabbit mAb #4058
Have you tried your application using our XP® monoclonal antibodies ? Try product: 4060
| Applications | Reactivity | Sensitivity | MW (kDa) | Isotype |
|---|---|---|---|---|
| W IP IF-IC F | H M R | Endogenous | 60 | Rabbit IgG |
Applications Key: W=Western Blotting IP=Immunoprecipitation IF-IC=Immunofluorescence (Immunocytochemistry) F=Flow Cytometry
Reactivity Key: H=Human M=Mouse R=Rat
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Protocols
- 4058:
- Flow , Immunofluorescence , Immunoprecipitation , Western Blotting
Specificity / Sensitivity
Phospho-Akt (Ser473) (193H12) Rabbit mAb detects endogenous levels of Akt only when phosphorylated at Ser473.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues around Ser473 of mouse Akt.
Western Blotting
Western blot analysis of extracts from untreated or PDGF-treated NIH/3T3 cells, pretreated with wortmannin and/or rapamycin as indicated, using Phospho-Akt (Ser473) (193H12) Rabbit mAb (upper) or Akt Antibody #9272 (lower).
Flow Cytometry
Flow cytometric analysis of Jurkat cells, untreated (green) or LY294002 and Wortmannin treated (blue), using Phospho-Akt (Ser473) (193H12) Rabbit mAb compared to a nonspecific negative control antibody (red).
IF-IC
Confocal immunofluorescent images of C2C12 cells either serum starved or treated with insulin as indicated and labeled with Phospho-Akt (Ser473) (193H12) Rabbit mAb (red). Actin filaments have been labeled with fluorescein phalloidin.
Background
Akt, also referred to as PKB or Rac, plays a critical role in controlling survival and apoptosis (1-3). This protein kinase is activated by insulin and various growth and survival factors to function in a wortmannin-sensitive pathway involving PI3 kinase (2,3). Akt is activated by phospholipid binding and activation loop phosphorylation at Thr308 by PDK1 (4) and by phosphorylation within the carboxy terminus at Ser473. The previously elusive PDK2 responsible for phosphorylation of Akt at Ser473 has been identified as mammalian target of rapamycin (mTOR) in a rapamycin-insensitive complex with rictor and Sin1 (5,6). Akt promotes cell survival by inhibiting apoptosis through phosphorylation and inactivation of several targets, including Bad (7), forkhead transcription factors (8), c-Raf (9), and caspase-9. PTEN phosphatase is a major negative regulator of the PI3 kinase/Akt signaling pathway (10). LY294002 is a specific PI3 kinase inhibitor (11). Another essential Akt function is the regulation of glycogen synthesis through phosphorylation and inactivation of GSK-3α and β (12,13). Akt may also play a role in insulin stimulation of glucose transport (12). In addition to its role in survival and glycogen synthesis, Akt is involved in cell cycle regulation by preventing GSK-3β-mediated phosphorylation and degradation of cyclin D1 (14) and by negatively regulating the cyclin dependent kinase inhibitors p27 Kip (15) and p21 Waf1/CIP1 (16). Akt also plays a critical role in cell growth by directly phosphorylating mTOR in a rapamycin-sensitive complex containing raptor (17). More importantly, Akt phosphorylates and inactivates tuberin (TSC2), an inhibitor of mTOR within the mTOR-raptor complex (18,19).
- Franke, T.F. et al. (1997) Cell 88, 435-7.
- Burgering, B.M. and Coffer, P.J. (1995) Nature 376, 599-602.
- Franke, T.F. et al. (1995) Cell 81, 727-36.
- Alessi, D.R. et al. (1996) EMBO J 15, 6541-51.
- Sarbassov, D.D. et al. (2005) Science 307, 1098-101.
- Jacinto, E. et al. (2006) Cell 127, 125-37.
- Cardone, M.H. et al. (1998) Science 282, 1318-21.
- Brunet, A. et al. (1999) Cell 96, 857-68.
- Zimmermann, S. and Moelling, K. (1999) Science 286, 1741-4.
- Cantley, L.C. and Neel, B.G. (1999) Proc Natl Acad Sci USA 96, 4240-5.
- Vlahos, C.J. et al. (1994) J Biol Chem 269, 5241-8.
- Hajduch, E. et al. (2001) FEBS Lett 492, 199-203.
- Cross, D.A. et al. (1995) Nature 378, 785-9.
- Diehl, J.A. et al. (1998) Genes Dev 12, 3499-511.
- Gesbert, F. et al. (2000) J Biol Chem 275, 39223-30.
- Zhou, B.P. et al. (2001) Nat Cell Biol 3, 245-52.
- Navé, B.T. et al. (1999) Biochem J 344 Pt 2, 427-31.
- Inoki, K. et al. (2002) Nat Cell Biol 4, 648-57.
- Manning, B.D. et al. (2002) Mol Cell 10, 151-62.
Application References
- Tu, Y. et al. (2007) J Biol Chem 282, 11722-31. Applications: Western Blotting
- Radhakrishnan, S. et al. (2007) J Immunol 178, 1426-32. Applications: Flow Cytometry Western Blotting
- Simoncic, P.D. et al. (2006) Mol Cell Biol 26, 4149-60. Applications: Western Blotting
- Wong, R.C. et al. (2007) Stem Cells Dev 16, 989-1001. Applications: Western Blotting
- Salvucci, O. et al. (2006) Blood 108, 2914-22. Applications: Flow Cytometry Western Blotting
- Berthou, S. et al. (2004) Oncogene 23, 5387-93. Applications: Western Blotting
- Sengupta, S. et al. (2005) Immunobiology 210, 647-59. Applications: Flow Cytometry
- Firaguay, G. and Nunès, J.A. (2009) Sci Signal 2, pl3. Applications: Flow Cytometry
- Benedettini, E. et al. (2010) Am J Pathol 177, 415-23. Applications: Western Blotting
- Popkie, A.P. et al. (2010) J Biol Chem 285, 41337-47. Applications: Western Blotting
- Billottet, C. et al. (2009) Cancer Res 69, 1027-36. Applications: Flow Cytometry
- Messal, N. et al. (2011) Eur J Immunol 41, 3443-54. Applications: Flow Cytometry
- Kleiman, L.B. et al. (2011) Mol Cell 43, 723-37. Applications: IF-IC (In Cells) Western Blotting
- Chung, Y.W. et al. (2011) J Biol Chem 286, 29681-90. Applications: IP Western Blotting
- Sykes, S.M. et al. (2011) Cell 146, 697-708. Applications: Western Blotting
- Dong, A. et al. (2011) J Biol Chem 286, 18301-10. Applications: Western Blotting
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know !
Companion Products
- 4060 Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb
- 2965 Phospho-Akt (Thr308) (C31E5E) Rabbit mAb
- 4691 Akt (pan) (C67E7) Rabbit mAb
- 4051 Phospho-Akt (Ser473) (587F11) Mouse mAb
- 3787 Phospho-Akt (Ser473) (736E11) Rabbit mAb
- 9272 Akt Antibody
- 2938 Akt1 (C73H10) Rabbit mAb
- 2964 Akt2 (5B5) Rabbit mAb
- 3788 Akt3 (62A8) Rabbit mAb
- 7160 PathScan® Phospho-Akt1 (Ser473) Sandwich ELISA Kit
- 7170 PathScan® Total Akt1 Sandwich ELISA Kit
- 9916 Phospho-Akt Pathway Antibody Sampler Kit
- 9901 LY294002 (PI3 Kinase Inhibitor)
- 9273 Akt Control Cell Extracts
- 7071 Phototope® -HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7003 20X LumiGLO® Reagent and 20X Peroxide
Rabbit Monoclonals Produced Using Epitomics® Technology, U.S. Patent No. 5,675,063.
For Research Use Only. Not For Use In Diagnostic Procedures.
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文献和实验(Ser473)(193H12)Rabbit mAb，1:300,稀释，二抗是驴抗兔，1：2000，我是实验室的新手，刚呆俩月，受 此挫折，一头雾水，请过来人或者有经验的同学帮忙解答怎样解决，非常感谢！ 碧峤 pAkt有两个位点，Ser473和Thr308，其中Ser473相对容易显色。如果显色失败，可能的原因:1 上样量过少，我们一般都是30μg；2 一抗孵育时间过短，我们一般24h~48h；3 一抗是否工作有待排除 米宝
：使用 Anti-phospho-Akt (Ser473) Rabbit mAb 对石蜡包埋的人乳腺癌组织进行免疫组织化学分析。（图 A）使用免疫组化试剂盒M&R HRP/DAB Detection IHC Kit，抗体 1:100 稀释；(图 B) 采用普通免疫组化试剂盒，抗体 1:25 稀释。 图 6 免疫组化实验检测 Erk1/2 表达 注：使用 Anti-Erk1/2 Mouse mAb与p44/42 MAPK （Erk1/2）Rabbit mAb 对正常小鼠心脏组织进行免疫
来实现的，抑制某种通路，可以理解为其磷酸化水平降低，具体可以通过免疫印迹看看其下游的蛋白表达是否降低了， 个人一点拙见，仅供参考 ylhuang0502 楼主提出一个很好的问题。 Y294002是常用的PI3K/AKT通路的抑制剂，如楼上所引，是通过对PI3K的竞争性抑制来抑制PI3K的酶活性，那么为什么大家常用phopho-AKT(S473)来检测呢，因为虽然众所周知Y294002可以PI3K的活性，但是没有一种磷酸化位点PI3K活性
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