Phospho-AKT (Ser473) Recombinant antibody

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

Host / Isotype

Rabbit / IgG

Reactivity

Human and More (3)

Applications

WB, IF, FC, ELISA

Conjugate

Unconjugated

Publications

20

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.

• 

  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.

• 

  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

Sample-dependent, check data in validation data gallery

Published Applications

• WB: See 19 publications below
• IF: See 1 publications below

Product Information

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

• Tested Reactivity: Human
• Cited Reactivity: human, mouse, rat
• 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
• 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	J Biomol Struct Dyn; Molecular dynamic simulation and functional analysis of pathogenic PTEN mutations in glioblastoma; Authors - Vildan Bozok Cetintas
human	WB	Pharmaceuticals (Basel); A Novel Mast Cell Stabilizer JM25-1 Rehabilitates Impaired Gut Barrier by Targeting the Corticotropin-Releasing Hormone Receptors; Authors - Yueshan Sun
rat	IF	Front Cell Dev Biol; Enhancing autophagy and energy metabolism in the meniscus can delay the occurrence of PTOA in ACLT rat; Authors - Huangrong Zhu
	WB	Ecotoxicol Environ Saf; Difenoconazole disrupts the blood-brain barrier and results in neurotoxicity in carp by inhibiting the Nrf2 pathway mediated ROS accumulation; Authors - Feixue Liu
rat	WB	Vascul Pharmacol; Midkine mediates dysfunction of liver sinusoidal endothelial cells through integrin α4 and α6; Authors - Li Wu
mouse	WB	J Cardiovasc Transl Res; Ferulic Acid Alleviates Atherosclerotic Plaques by Inhibiting VSMC Proliferation Through the NO/p21 Signaling pathway.; Authors - Xiaoyun Wu