- ￥1500 - 3280
- Proteintech
- 66444-1-Ig
- 2025年10月13日
- WB, IHC, IF/ICC, FC (Intra), ELISA
- Mouse
- human, mouse, rat
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- 详细信息
- 询价记录
- 文献和实验
- 技术资料
- 免疫原：
None
- 亚型：
IgG1
- 保存条件：
Store at -20°C. Stable for one year after shipment.
- 克隆性：
Monoclonal
- 标记物：
Unconjugated
- 适应物种：
human, mouse, rat
- 保质期：
详见产品包装
- 库存：
充足
- 宿主：
Mouse
- 应用范围：
WB, IHC, IF/ICC, FC (Intra), ELISA
- 靶点：
Phospho-AKT (Ser473)
- 抗体英文名：
Phospho-AKT (Ser473) Monoclonal antibody
- 抗体名：
Phospho-AKT (Ser473) Monoclonal antibody
- 规格：
50ul/100ul/150ul
| 规格： | 50ul | 产品价格： | ￥1500.0 |
|---|---|---|---|
| 规格： | 100ul | 产品价格： | ￥2480.0 |
| 规格： | 150ul | 产品价格： | ￥3280.0 |
经过测试的应用
| Positive WB detected in | Calyculin A treated PC-3 cells, Calyculin A treated HEK-293T cells, HSC-T6 cells, TPA treated Jurkat cells, Calyculin A treated HSC-T6 cells |
| Positive IHC detected in | human breast cancer tissue, Calyculin A treated Jurkat cells, human colon cancer tissue Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0 |
| Positive IF/ICC detected in | Calyculin A treated HeLa cells |
| Positive FC (Intra) detected in | Calyculin A treated PC-3 cells |
推荐稀释比
| 应用 | 推荐稀释比 |
|---|---|
| Western Blot (WB) | WB : 1:2000-1:10000 |
| Immunohistochemistry (IHC) | IHC : 1:100-1:400 |
| Immunofluorescence (IF)/ICC | IF/ICC : 1:200-1:800 |
| Flow Cytometry (FC) (INTRA) | FC (INTRA) : 0.50 ug per 10^6 cells in a 100 µl suspension |
| It is recommended that this reagent should be titrated in each testing system to obtain optimal results. | |
| Sample-dependent, Check data in validation data gallery. | |
产品信息
66444-1-Ig targets Phospho-AKT (Ser473) in WB, IHC, IF/ICC, FC (Intra), ELISA applications and shows reactivity with human, mouse, rat samples.
| 经测试应用 | WB, IHC, IF/ICC, FC (Intra), ELISA |
| 文献引用应用 | WB, IHC, IF |
| 经测试反应性 | human, mouse, rat |
| 文献引用反应性 | human, mouse, rat, pig, rabbit, monkey, chicken, zebrafish, sheep, duck |
| 免疫原 |
fusion protein |
| 宿主/亚型 | Mouse / IgG1 |
| 抗体类别 | Monoclonal |
| 产品类型 | Antibody |
| 全称 | v-akt murine thymoma viral oncogene homolog 1 |
| 别名 | AKT (Ser473), P AKT, P AKT (Ser473), p AKT Ser473, P-AKT |
| 观测分子量 | 60-62 kDa |
| GenBank蛋白编号 | NM_005163 |
| 基因名称 | AKT1 |
| Gene ID (NCBI) | 207 |
| RRID | AB_2782958 |
| 偶联类型 | Unconjugated |
| 形式 | Liquid |
| 纯化方式 | Protein G purification |
| UNIPROT ID | P31749 |
| 储存缓冲液 | PBS with 0.02% sodium azide and 50% glycerol, pH 7.3. |
| 储存条件 | Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20oC storage. |
背景介绍
1) What is AKT?
The serine/threonine kinase B AKT pathway (also known as the PI3K-Akt pathway) plays a vital role in the regulation of cellular processes, including cell proliferation, survival, and growth - processes that are essential for oncogenesis. Mutation of the regulator proteins PI3K and PTEN causes uncontrolled disruption within the PI3-kinase pathway, leading to the development of human cancers (1,2; see also AKT pathway poster for more details).
2) phospho-AKT and FAQs
A) What is the best way to normalize phosphorylated proteins analyzed by western blot?
Normalize phospho-AKT and total AKT with your loading control (e.g. Actin, tubulin), then calculate the phospho/total ratio using these normalized values.
Put more simply:
1. Calculate the ratio of band intensities of a phospho-AKT band: the loading control.
2. Calculate the ratio of band intensities of total AKT: loading control.
3. Divide ratio obtained #1 by #2 to obtain a normalized value for comparison among different conditions. This procedure allows one to distinguish between a change in AKT expression and a change in the ratio of phospho-AKT.
* If you are looking at the differences in a phospho-AKT expression resulting from an experimental condition (e.g., knockdown), you should also show the expression of total AKT to distinguish between a change in AKT expression (transcription/translation level) and a change in the AKT phosphorylation status.
B) What is the observed molecular weight for AKT and phospho-AKT?
Molecular Weight AKT - 56 kDa
Molecular Weight phospho-AKT - 60 kDa (Figure 1)
Figure 1. WB: HEK-293 cell lysate was subjected to SDS PAGE followed by western blot with 60203-2-Ig (AKT antibody) and 66444-1-Ig (AKT-phospho-S473 antibody) at a dilution of 1:4000 incubated at room temperature for 1.5 hours.
C) Are there any special WB conditions to optimize staining of a phospho-AKT?
Since this is a phosphorylated protein, 5% BSA is recommended over non-fat milk as a blocking agent.
D) What are good positive and negative controls for a phospho-AKT?
- Positive Control: HEK293 cells
- Negative Control: Treatment with PI3K inhibitors (e.g. wortmannin)
E) What species does this antibody react with?
Our internal testing has confirmed that it reacts with the human and mouse forms of phospho-AKT.Reactivity with the human form is also supported by the literature's citations of this antibody.
References:
1. Perturbations of the AKT signaling pathway in human cancer.
2. Targeting the PI3K-Akt pathway in human cancer: rationale and promise.
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- 作者
- 内容
- 询问日期
文献和实验| Species | Application | Title |
|---|---|---|
| human | WB | Signal Transduct Target Ther Circulating tumor cells shielded with extracellular vesicle-derived CD45 evade T cell attack to enable metastasisAuthors - Chuan Yang |
Adv Mater Targeted Macrophage CRISPR-Cas13 Mrna Editing in Immunotherapy for Tendon InjuryAuthors - Shuo Wang | ||
| mouse | WB | Cell Metab Disrupted methionine cycle triggers muscle atrophy in cancer cachexia through epigenetic regulation of REDD1Authors - Kai Lin |
| mouse | WB,IHC | Cell Res In vivo self-assembled small RNAs as a new generation of RNAi therapeutics.Authors - Zheng Fu |
| mouse | IF | Cell Res Inhibiting Hv1 channel in peripheral sensory neurons attenuates chronic inflammatory pain and opioid side effects.Authors - Qiansen Zhang |
| mouse,human | WB | Nat Commun Genome-wide enhancer-gene regulatory maps link causal variants to target genes underlying human cancer riskAuthors - Pingting Ying |
出来的.像前面几位战友说的那样,可以考虑加大上样量 2. 是否WB的条件不好? 可以跑其他蛋白的WB看看,如果其他蛋白质都跑出来的很好的话, 可以考虑换个抗体试一试,,但是话说回来,cell signaling的p-AKT 很好的,我也用了好几年.ser473和T308都不错的. 3. 条带弱的话,可以用 can get sinal 去提高抗体的敏感度. BTW, 我自己跑p-akt的时候, 1抗是1:1500 2抗是 1:20000 good luck
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
receptorr 我在做HaCaT细胞的磷酸化Akt(473)时，背景比较高，该怎么办？（即未处理组的Akt比较高） 请高手支招。 谢谢 magichunter 说得太粗略了。 receptorr 我做的细胞是HaCaT 看看加入药物后HaCaT的磷酸化Akt是否有增高， 所以希望没有加药的那一组HaCaT的磷酸化Akt水平低 但是实际做时，发现没有
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