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- 详细信息
- 文献和实验
- 技术资料
- 抗体英文名:
Akt (pan) (11E7) Rabbit mAb
- 抗原:
synthetic peptide at the carboxy-terminal sequence of mouse Akt
- 应用范围:
W, IP, IHC-P, IF-IC, F
- 供应商:
CST
- 库存:
大量
- 级别:
详见MSDS文件
- 适应物种:
H,M,R,Mk
- 保质期:
详见说明书
- 是否单克隆:
单克隆
- 保存条件:
-20°c
pathway more info application references datasheet PDF MSDS PDF protocols
Applications Key: W=Western Blotting IP=Immunoprecipitation IHC-P=Immunohistochemistry (Paraffin) IF-IC=Immunofluorescence (Immunocytochemistry) F=Flow Cytometry
Reactivity Key: H=Human M=Mouse R=Rat Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
| Applications | Reactivity | Sensitivity | MW (kDa) | Isotype |
|---|---|---|---|---|
| W IP IHC-P IF-IC F | H M R Mk | Endogenous | 60 | Rabbit IgG |
| Protocols |
|
|---|---|
| Specificity / Sensitivity | Akt (pan) (11E7) Rabbit mAb detects endogenous levels of total Akt protein. This antibody does not cross-react with other related proteins. |
| Source / Purification | Monoclonal antibody is produced by immunizing animals with a synthetic peptide at the carboxy-terminal sequence of mouse Akt. Western Blotting
Western blot analysis of recombinant Akt1, Akt2 and Akt3 proteins, and extracts from HeLa, C2C12, C6 and COS cells, using Akt (pan) (11E7) Rabbit mAb. IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Akt (pan) (11E7) Rabbit mAb in the presence of control peptide (left) or Akt (pan) (11E7) Blocking Peptide #1085 (right). IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using Akt (pan) (11E7) Rabbit mAb. IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human melanoma, using Akt (11E7) Rabbit mAb. IHC-P (paraffin)
Immunohistochemical analysis using Akt (pan) (11E7) Rabbit mAb on SignalSlide(TM) Phospho-Akt (Ser473) IHC Controls #8101 (paraffin-embedded LNCaP cells, untreated (left) or LY294002-treated (right)). Note the lack of phosphorylated Akt-associated stain at the membrane of the LY294002 treated cells. IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded HeLa cells untreated (left) or transfected with Akt 1/2/3 Kinases ShortCut® siRNA Mix (New England BioLabs #N2005) (right), using Akt (pan) (11E7) Rabbit mAb (top) or Cleaved Caspase-3 (Asp175) #9661 (bottom). Note the induction of cleaved caspase-3 in Akt deficient cells. |
| 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).
|
| Application References |
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 |
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
组织化学分析。(图 C)使用Anti-Erk1/2 Mouse mAb 鼠单抗进行目的蛋白检测;(图 D) 使用p44/42 MAPK (Erk1/2)Rabbit mAb 兔单抗进行目的蛋白检测。 图 7 免疫组化实验检测Akt表达 注:使用 Anti-Akt (pan) Rabbit mAb 对正常小鼠肝组织进行免疫组织化学分析。(图 E)使用免疫组化试剂盒 M&R HRP/DAB Detection IHC Kit,抗体1:100 稀释;(图F)使用另一种试剂盒,抗体 1:100 稀释。
过程中,一抗应在封闭缓冲液 (TBS/0.3%Triton. X–100/5% NGS) 中稀释。抗体:Phospho–Akt (Ser473) (D9E) XP. Rabbit mAb #4060、Phospho–EGFReceptor (Tyr1173) (53A5) Rabbit mAb #4407样本:石蜡包埋人类乳腺肿瘤(上图)和 HCC827 异种移植物(下图)抗体稀释:SignalStain. 抗体稀释液(左)或 TBST/5%NGS(右)检测—检测系统传统的 IHC 检测
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