- ￥2800
- Proteintech
- CL555-66444
- 2025年10月09日
- FC (Intra)
- Mouse
- human, mouse
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- 详细信息
- 文献和实验
- 技术资料
- 亚型：
IgG1
- 保存条件：
Store at -20°C. Avoid exposure to light. Stable for one year after shipment.
- 克隆性：
Monoclonal
- 标记物：
CoraLite®555 Fluorescent Dye
- 适应物种：
human, mouse
- 宿主：
Mouse
- 应用范围：
FC (Intra)
- 靶点：
Phospho-AKT (Ser473)
- 抗体英文名：
CoraLite®555-conjugated Phospho-AKT (Ser473) Monoclonal antibody
- 抗体名：
CoraLite®555-conjugated Phospho-AKT (Ser473) Monoclonal antibody
- 规格：
100ul
经过测试的应用
| Positive FC (Intra) detected in | Calyculin A treated PC-3 cells |
推荐稀释比
| 应用 | 推荐稀释比 |
|---|---|
| Flow Cytometry (FC) (INTRA) | FC (INTRA) : 0.13 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
CL555-66444 targets Phospho-AKT (Ser473) in FC (Intra) applications and shows reactivity with human, mouse samples.
| 经测试应用 | FC (Intra) |
| 经测试反应性 | human, mouse |
| 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 pan, AKT 1, AKT (pan), AKT, AKT1 |
| 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_2934673 |
| 偶联类型 | CoraLite®555 Fluorescent Dye |
| 最大激发/发射波长 | 557 nm / 570 nm |
| 形式 | Liquid |
| 纯化方式 | Protein A purification |
| UNIPROT ID | P31749 |
| 储存缓冲液 | PBS with 50% glycerol, 0.05% Proclin300, 0.5% BSA, pH 7.3. |
| 储存条件 | Store at -20°C. Avoid exposure to light. 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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文献和实验过程中，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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SAB(Signalway antibody ) 是一家汇聚了多名专家和学者的新型高科技生物公司，其研发和生产的抗体包括了数百种磷酸化 / 非磷酸化抗体 ( 可以提供同一位点的磷酸化 / 非磷酸化的成对抗体 ) ，为神经科学、细胞信号转导 (Akt pathway ， Jat/Sat pathway ， MAPK pathway ， NF-Kappa B pathway 等 ) 、癌症 / 凋亡、细胞周期、染色质 / 转录、细胞骨架通道、胰岛素 / 葡萄糖、核受体 / 膜受体
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