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- 详细信息
- 文献和实验
- 技术资料
- 供应商:
上海联迈生物工程有限公司
- 库存:
大量
- 目录编号:
LM-2720R
- 克隆性:
多克隆
- 抗原来源:
Rabbit
- 保质期:
1年
- 抗体英文名:
phospho-Akt (Thr308)
- 抗体名:
磷酸化蛋白激酶B抗体
- 宿主:
Rabbit
- 适应物种:
Human, Mouse, Rat, Chicken, Dog, Pig, Cow, Rabbit, Sheep,
- 免疫原:
KLH conjugated Synthesised phosphopeptide derived from human Akt around the phosphorylation site of Thr308:MK(p-T)FC
- 亚型:
IgG
- 形态:
Lyophilized or Liquid
- 应用范围:
WB=1:500-2000 ELISA=1:500-1000 IHC-P=1:400-800 IHC-F=1:400-800 Flow-Cyt=1μg /test IF=1:100-500 (石蜡切片需做抗原修复)
- 浓度:
1mg/ml
- 保存条件:
Store at -20 °C
- 规格:
100ul
| 英文名称 | phospho-Akt (Thr308) |
| 中文名称 | 磷酸化蛋白激酶B抗体 |
| 别 名 | AKT (phospho T308); p-AKT (phospho T308); Akt(Phospho-Thr308); Akt(Phospho T308); C AKT; MGC9965; Oncogene AKT1; PKB; PRKBA; Protein Kinase B Alpha; RAC Alpha; RAC; RAC PK Alpha; RAC Serine/Threonine Protein Kinase; vAKT Murine Thymoma Viral Oncogene Homolog 1; AKT1_HUMAN. |
| 规格价格 | 100ul/1580元 购买 大包装/询价 |
| 说 明 书 | 100ul |
| 产品类型 | 磷酸化抗体 |
| 研究领域 | 细胞生物 神经生物学 信号转导 细胞凋亡 细胞周期蛋白 激酶和磷酸酶 |
| 抗体来源 | Rabbit |
| 克隆类型 | Polyclonal |
| 交叉反应 | Human, Mouse, Rat, Chicken, Dog, Pig, Cow, Rabbit, Sheep, |
| 产品应用 | WB=1:500-2000 ELISA=1:500-1000 IHC-P=1:400-800 IHC-F=1:400-800 Flow-Cyt=1μg /test IF=1:100-500 (石蜡切片需做抗原修复) not yet tested in other applications. optimal dilutions/concentrations should be determined by the end user. |
| 分 子 量 | 56kDa |
| 细胞定位 | 细胞核 细胞浆 细胞膜 |
| 性 状 | Lyophilized or Liquid |
| 浓 度 | 1mg/ml |
| 免 疫 原 | KLH conjugated Synthesised phosphopeptide derived from human Akt around the phosphorylation site of Thr308:MK(p-T)FC |
| 亚 型 | IgG |
| 纯化方法 | affinity purified by Protein A |
| 储 存 液 | 0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol. |
| 保存条件 | Store at -20 °C for one year. Avoid repeated freeze/thaw cycles. The lyophilized antibody is stable at room temperature for at least one month and for greater than a year when kept at -20°C. When reconstituted in sterile pH 7.4 0.01M PBS or diluent of antibody the antibody is stable for at least two weeks at 2-4 °C. |
| PubMed | PubMed |
| 产品介绍 | background: The serine-threonine protein kinase encoded by the AKT1 gene is catalytically inactive in serum-starved primary and immortalized fibroblasts. AKT1 and the related AKT2 are activated by platelet-derived growth factor. The activation is rapid and specific, and it is abrogated by mutations in the pleckstrin homology domain of AKT1. It was shown that the activation occurs through phosphatidylinositol 3-kinase. In the developing nervous system AKT is a critical mediator of growth factor-induced neuronal survival. Survival factors can suppress apoptosis in a transcription-independent manner by activating the serine/threonine kinase AKT1, which then phosphorylates and inactivates components of the apoptotic machinery. Mutations in this gene have been associated with the Proteus syndrome. Multiple alternatively spliced transcript variants have been found for this gene. [provided by RefSeq, Jul 2011] Subunit: Interacts (via PH domain) with TCL1A; this enhances AKT3 phosphorylation and activation. Interacts with TRAF6. Subcellular Location: Cytoplasm. Nucleus. Cell membrane. Note=Nucleus after activation by integrin-linked protein kinase 1 (ILK1). Nuclear translocation is enhanced by interaction with TCL1A. Phosphorylation on Tyr-176 by TNK2 results in its localization to the cell membrane where it is targeted for further phosphorylations on Thr-308 and Ser-473 leading to its activation and the activated form translocates to the nucleus. Tissue Specificity: Widely expressed. Low levels found in liver with slightly higher levels present in thymus and testis. Similarity: Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. RAC subfamily. Contains 1 AGC-kinase C-terminal domain. Contains 1 PH domain. Contains 1 protein kinase domain. SWISS: P31749 Gene ID: 207 Database links: Entrez Gene: 207 Human Entrez Gene: 11651 Mouse Entrez Gene: 24185 Rat Omim: 164730 Human SwissProt: O57513 Chicken SwissProt: P31749 Human SwissProt: P31750 Mouse SwissProt: P47196 Rat Unigene: 525622 Human Unigene: 6645 Mouse Unigene: 11422 Rat Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications. |
| 产品图片 | ![]() Sample: Liver (Mouse) Lysate at 30 ug Primary: Anti- phospho-Akt(Thr308)(bs-2720R) at 1/300 dilution Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/10000 dilution Predicted band size: 56 kD Observed band size: 56 kD ![]() Western blot analysis of extracts from Heart,using phospho-Akt(thr308) Antibody. ![]() Paraformaldehyde-fixed, paraffin embedded (Mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (phospho-Akt (Thr308)) Polyclonal Antibody, Unconjugated (bs-2720R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructions and DAB staining. ![]() Paraformaldehyde-fixed, paraffin embedded (Rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (phospho-Akt (Thr308)) Polyclonal Antibody, Unconjugated (bs-2720R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructions and DAB staining. ![]() Blank control (blue line): A431(Black). Primary Antibody (green line): Rabbit Anti-phospho-Akt (Thr308) antibody (bs-2720R) Dilution: 1μg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG . Secondary Antibody (white blue line): Goat anti-rabbit IgG-PE(Jackson lab) Dilution: 1μg /test. Protocol The cells were fixed with 4% paraformaldehyde (10 min) , then permeabilized with 90% ice-cold methanol for 20 min on ice. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed. |
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文献和实验)in TBST 室温一个小时,一抗稀释在5%BSA in TBST 中(而非5%脱脂牛奶)4度过夜,这样我相信背景会降低,提高信噪比。 补充一些, 理论上, 用WB测蛋白的磷酸化的时候,常规是BSA比牛奶更好. 因为牛奶中含有casein, 会非特异性的和测磷酸化的抗体结合,导致背景着色提高.AKT的磷酸化,我自己也用过BSA 也用过脱脂奶粉.感觉没多大区别. 磷酸化如果测不出来的时候 1. 可能是这个组织或细胞本身的磷酸化很低. 但是你用的那几个脏器,比如肝肺 都能测
prince1101 非磷酸化抗体是识别总得蛋白的,比如AKT。 P-AKT和AKT其实只差一些磷酸基,分子量稍大点,以前听别人讲P-AKT和AKT之间有shift,有时候WB是可以做出来的。 用的是梯度胶?分子量在68KD左右。应该怎么样来配胶呢?梯度胶的浓度? 谢谢! prince1101 希望高手能赐教! ourlab 某些 AKT抗体,如cell
米宝 这俩天做实验,极其郁闷。我做的总AKt和pAkt,大鼠脑组织,第一次跑总AKT的时候,我觉得可能是因为蛋白是提取的浓度比较好吧,出来了一条浓浓的条带,三组总AKt都出了,接下来做pAkt,就出了一个带,效果挺明显的。后来再做pAkt的时候,跑了4张膜,显影的时候,太干净了,上面什么都没有,甚至连个杂带,斑点 都没!实验是在实验员的领导下完成的,技术上基本没什么纰漏,查阅本版面有关这问题方面的介绍,有的解释说pakt降解了!一抗是用的Phospho-Akt
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