Phospho-AKT (Ser473) Recombinant antibody

Phospho-AKT (Ser473) Recombinant Antibody for FC, WB, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

Human and More (4)

Applications

WB,FC,ELISA

Conjugate

Unconjugated

Publications

51

CloneNo.

2E17

Cat no : 80455-1-RR

Synonyms

AKT, Phospho-AKT (Ser473), PKB, PKB ALPHA, PRKBA, Protein kinase B, Proto oncogene c Akt, RAC, RAC ALPHA, RAC PK alpha

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Validation Data Gallery

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  WB analysis of HEK-293 using 80455-1-RR

  Non-treated and Calyculin A treated HEK-293 cells were subjected to SDS PAGE followed by western blot with 80455-1-RR (Phospho-AKT (Ser473) antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with GAPDH antibody as loading control.

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  WB analysis of HeLa using 80455-1-RR

  Non-treated and Calyculin A treated HeLa cells were subjected to SDS PAGE followed by western blot with 80455-1-RR (Phospho-AKT (Ser473) antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with GAPDH antibody as loading control.

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  WB analysis of HEK-293T using 80455-1-RR

  Non-treated and IGF-1 treated HEK-293T cells were subjected to SDS PAGE followed by western blot with 80455-1-RR (Phospho-AKT (Ser473) antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with GAPDH antibody as loading control.

• 

  FC experiment of HEK-293 using 80455-1-RR

  1X10^6 HEK-293 cells untreated (dashed lines) or treated with Calyculin A (red) were intracellularly stained with 0.25 ug Anti-Human Phospho-AKT (Ser473) (80455-1-RR, Clone:2E17) and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) at dilution 1:1000, or 0.25 ug Control Antibody (blue). Cells were fixed with 4% PFA and permeabilized with 80% MeOH.

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Tested Applications

• Positive WB detected in: HEK-293 cells, HEK-293T cells, HeLa cells, IGF-1 treated HEK-293T cells, Calyculin A treated HEK-293 cells, Calyculin A treated HeLa cells
• Positive FC detected in: Calyculin A treated HEK-293 cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Flow Cytometry (FC): FC : 0.25 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.

Published Applications

• WB: See 50 publications below
• IHC: See 1 publications below
• IF: See 1 publications below

Product Information

80455-1-RR targets Phospho-AKT (Ser473) in WB,FC,ELISA applications and shows reactivity with Human samples.

• Tested Reactivity: Human
• Cited Reactivity: human, mouse, rat, bovine
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: Peptide
• Full Name: v-akt murine thymoma viral oncogene homolog 1
• Observed Molecular Weight: 58 kDa
• GenBank Accession Number: NM_005163
• Gene Symbol: AKT1
• Gene ID (NCBI): 207
• RRID: AB_2918892
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

AKT is a serine/threonine kinase and it participates in the key role of the PI3K signaling pathway. Phosphatidylinositol-3 kinase (PI3K) is the key regulator of AKT activation. The recruitment of inactive AKT protein to PIP3-rich areas of the plasma membrane results in a conformational change that exposes the activation loop of AKT. AKT's activating kinase, phosphoinositide-dependent protein kinase (PDK1) , is also recruited to PIP3 microdomains. PDK1 phosphorylates AKT on threonine 308 (Thr308) of the exposed activation loop, activating AKT and leading to a second phosphorylation of AKT at serine 473 (Ser473) by a kinase presumed to be mTORC2 that further potentiates kinase activity. Active AKT will phosphorylate various downstream protein targets that control cell growth and translational control and act to suppress apoptosis. (PMID: 31594388, PMID: 30808672)

Publications

Species	Application	Title
human	WB	Adv Healthc Mater; A ROS-Responsive Liposomal Composite Hydrogel Integrating Improved Mitochondrial Function and Pro-Angiogenesis for Efficient Treatment of Myocardial Infarction.; Authors - Zhi Zheng
mouse	WB	Research (Wash D C); Herpetrione, a New Type of PPARα Ligand as a Therapeutic Strategy Against Nonalcoholic Steatohepatitis; Authors - Lang Linghu
human	WB	Arch Pharm Res; Selumetinib overcomes gefitinib primary and acquired resistance by regulating MIG6/STAT3 in NSCLC; Authors - Xiaoping Song
human	WB	iScience; WIPF1 promotes gastric cancer progression by regulating PI3K/Akt signaling in a myocardin-dependent manner; Authors - Fei Su
rat	WB	Int J Mol Sci; Identification of Potential Mechanisms of Rk1 Combination with Rg5 in the Treatment of Type II Diabetes Mellitus by Integrating Network Pharmacology and Experimental Validation; Authors - Yao Liu
human	WB	J Cell Mol Med; Irreversible repolarization of tumour-associated macrophages by low-Pi stress inhibits the progression of hepatocellular carcinoma; Authors - Yang-Feng Lv