重组Anti-AKT1 (pS473) + AKT2 (pS474) + AKT3 (pS472)抗体[EPR18853] - BSA and Azide free (ab222489)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18853] to AKT3 (phospho S472) + AKT2 (phospho S474) + AKT1 (phospho S473) - BSA and Azide free
- Suitable for: WB, ICC/IF, IP, Dot blot
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-AKT1 (pS473) + AKT2 (pS474) + AKT3 (pS472)抗体[EPR18853] - BSA and Azide free -
描述
兔单克隆抗体[EPR18853] to AKT3 (phospho S472) + AKT2 (phospho S474) + AKT1 (phospho S473) - BSA and Azide free -
宿主
Rabbit -
经测试应用
适用于: WB, ICC/IF, IP, Dot blotmore details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: MCF7 whole cell lysate treated with 100ng/ml IGF-1 for 15 minutes; PC-12 and NIH/3T3 whole cell lysates treated with 100ng/ml PDGF for 60 minutes. ICC/IF: NIH/3T3 cells treated with PDGF (100 ng/ml) for 1 hour. IP: NIH/3T3 treated with 50ng/ml PDGF for 40min whole cell lysate.
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常规说明
ab222489 is the carrier-free version of ab192623.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
存储溶液
pH: 7.2
Constituent: PBS -
无载体
是 -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR18853 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab222489于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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WB | (1) |
Use at an assay dependent concentration. Detects a band of approximately 56 kDa (predicted molecular weight: 56 kDa).
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ICC/IF |
Use at an assay dependent concentration.
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IP |
Use at an assay dependent concentration.
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Dot blot |
Use at an assay dependent concentration.
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说明 |
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WB
Use at an assay dependent concentration. Detects a band of approximately 56 kDa (predicted molecular weight: 56 kDa). |
ICC/IF
Use at an assay dependent concentration. |
IP
Use at an assay dependent concentration. |
Dot blot
Use at an assay dependent concentration. |
靶标
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细胞定位
AKT3: Cytoplasm. Membrane. Membrane-associated after cell stimulation leading to its translocation. AKT1: Cytoplasm. Nucleus. Cell membrane. 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. -
数据库链接
- Entrez Gene: 10000 Human
- Entrez Gene: 207 Human
- Entrez Gene: 208 Human
- Entrez Gene: 11651 Mouse
- Entrez Gene: 11652 Mouse
- Entrez Gene: 23797 Mouse
- Entrez Gene: 24185 Rat
- Entrez Gene: 25233 Rat
see all
图片
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells, untreated or treated with PDGF (100 ng/ml) for 1 hour, labeling AKT3 (phospho S472) + AKT2 (phospho S474) + AKT1 (phospho S473) with ab192623 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
The signal increased after treatment with PDGF (100 ng/ml) for 1 hour on NIH/3T3 cells.
The nuclear counter stain is DAPI (blue).
Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab192623).
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AKT3 (phospho S472) was immunoprecipitated from 0.35 mg of NIH/3T3 (Mouse embryonic fibroblast cell line) treated with 50ng/ml PDGF for 40min whole cell lysate with ab192623 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab192623 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.
Lane 1: NIH/3T3 treated with 50ng/ml PDGF for 40min whole cell lysate, 10μg (Input).
Lane 2: ab192623 IP in NIH/3T3 treated with 50ng/ml PDGF for 40min whole cell lysate.
Lane 3: Rabbit IgG,monoclonal [EPR25A]-Isotype Control (ab172730) instead of ab192623 in NIH/3T3 treated with 50ng/ml PDGF for 40min whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 5 seconds.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab192623).
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Dot blot analysis of AKT3 (phospho S472) labeled with ab192623 at 1/1000 dilution.
Lane 1: AKT3 (phospho S472) phospho peptide;
Lane 2: AKT3 non-phospho peptide;
Lane 3: AKT1 (phospho S473) phospho peptide;
Lane 4: AKT2 (phospho S474) phospho peptide.
Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated (ab97051) at 1/100000 dilution was used as secondary antibody.
Blocking and diluting buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab192623).
实验方案
数据表及文件
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Datasheet download
Certificate of Compliance
文献 (2)
ab222489 被引用在 2 文献中.
- Chen Y et al. Dendrobium mixture attenuates renal damage in rats with diabetic nephropathy by inhibiting the PI3K/Akt/mTOR pathway. Mol Med Rep 24:N/A (2021). PubMed: 34165163
- Sheng H et al. Combined Pharmacotherapy with Alendronate and Desferoxamine Regulate the Bone Resorption and Bone Regeneration for Preventing Glucocorticoids-Induced Osteonecrosis of the Femoral Head. Biomed Res Int 2020:3120458 (2020). PubMed: 33029500