重组Anti-AKT1 (pS473) + AKT2 (pS474) + AKT3 (pS472)抗体[EPR18853] (ab192623)
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)
- Suitable for: ICC/IF, WB, 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] -
描述
兔单克隆抗体[EPR18853] to AKT3 (phospho S472) + AKT2 (phospho S474) + AKT1 (phospho S473) -
宿主
Rabbit -
经测试应用
适用于: ICC/IF, WB, 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
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 short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
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ab192623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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ICC/IF |
1/100.
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WB |
1/1000. Detects a band of approximately 56 kDa (predicted molecular weight: 56 kDa).
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IP |
1/40.
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Dot blot |
1/1000.
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说明 |
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ICC/IF
1/100. |
WB
1/1000. Detects a band of approximately 56 kDa (predicted molecular weight: 56 kDa). |
IP
1/40. |
Dot blot
1/1000. |
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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All lanes : Anti-AKT1 (pS473) + AKT2 (pS474) + AKT3 (pS472) antibody [EPR18853] (ab192623) at 1/1000 dilution
Lane 1 : LNCaP (Human prostate carcinoma epithelial cell) whole cell lysates
Lane 2 : LNCaP (Human prostate carcinoma epithelial cell) treated with 0.1 uM Calyculin A for 30 minutes whole cell lysates
Lane 3 : A549 (Human lung carcinoma epithelial cell) whole cell lysates
Lane 4 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysates
Lane 5 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates
Lane 6 : HUVEC (Human umbilical vein endothelial cell) whole cell lysates
Lane 7 : C2C12 (Mouse myoblasts myoblast) whole cell lysates
Lane 8 : Mouse brain lysates
Lane 9 : Rat heart lysates
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 56 kDa
Observed band size: 56 kDa
Exposure time: 50 secondsThe basal expression level of AKT1 (phospho S473) varies in different cell lines reported by PMID: 19372546. But to detect clear signal, treatment is strongly recommended when using this antibody.
Blocking and Diluting buffer: 5% NFDM/TBST
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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.
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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.
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All lanes : Anti-AKT1 (pS473) + AKT2 (pS474) + AKT3 (pS472) antibody [EPR18853] (ab192623) at 1/1000 dilution
Lane 1 : Untreated MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
Lane 2 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate treated with 100ng/ml IGF-1 for 15 minutes
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 56 kDa
Observed band size: 56 kDa
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
Phosphorylation of AKT at S473 can be induced by IGF-1 treatment according to the literature (PMID: 23638184).
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All lanes : Anti-AKT1 (pS473) + AKT2 (pS474) + AKT3 (pS472) antibody [EPR18853] (ab192623) at 1/1000 dilution
Lane 1 : Untreated PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate treated with 100ng/ml PDGF for 60 minutes
Lane 3 : Untreated NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
Lane 4 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate treated with 100ng/ml PDGF for 60 minutes
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 56 kDa
Observed band size: 56 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1 and 2: 15 seconds; Lane 3 and 4: 3 minutes.
Phosphorylation of AKT can be induced by PDGF treatment according to the literature (PMID: 10984605 and 7774014).
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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.
实验方案
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (53)
ab192623 被引用在 53 文献中.
- Wu CT et al. SARS-CoV-2 replication in airway epithelia requires motile cilia and microvillar reprogramming. Cell 186:112-130.e20 (2023). PubMed: 36580912
- Kotulova J et al. 2‑Cl‑IB‑MECA regulates the proliferative and drug resistance pathways, and facilitates chemosensitivity in pancreatic and liver cancer cell lines. Int J Mol Med 49:N/A (2022). PubMed: 35039871
- Zhang G et al. Radix Astragalus Polysaccharide Accelerates Angiogenesis by Activating AKT/eNOS to Promote Nerve Regeneration and Functional Recovery. Front Pharmacol 13:838647 (2022). PubMed: 35431954
- Shibata O et al. Establishment of a pancreatic cancer animal model using the pancreas-targeted hydrodynamic gene delivery method. Mol Ther Nucleic Acids 28:342-352 (2022). PubMed: 35474735
- Zhou Y et al. Overexpression of microRNA-145 enhanced docetaxel sensitivity in breast cancer cells via inactivation of protein kinase B gamma-mediated phosphoinositide 3-kinase -protein kinase B pathway. Bioengineered 13:11310-11320 (2022). PubMed: 35499128