Phospho-AKT1 (Ser473) Recombinant antibody

Phospho-AKT1 (Ser473) Recombinant Antibody for FC, WB, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

Human, mouse and More (1)

Applications

WB, FC, ELISA

Conjugate

Unconjugated

Publications

2

CloneNo.

2M10

Cat no : 80462-1-RR

Synonyms

AKT, AKT1, Phospho-AKT (Ser473), Phospho-AKT1 (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 using 80462-1-RR

  Non-treated and Calyculin A treated HeLa cells were subjected to SDS PAGE followed by western blot with 80462-1-RR (Phospho-AKT1 (Ser473) antibody) at dilution of 1:5000 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 using 80462-1-RR

  Non-treated and Calyculin A treated NIH/3T3 cells were subjected to SDS PAGE followed by western blot with 80462-1-RR (Phospho-AKT1 (Ser473) antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with GAPDH antibody as loading control.

• 

  WB analysis using 80462-1-RR

  Non-treated and IGF1 treated HEK-293T cells were subjected to SDS PAGE followed by western blot with 80462-1-RR (Phospho-AKT1 (Ser473) antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with GAPDH antibody as loading control.

• 

  WB analysis using 80462-1-RR

  Non-treated and Calyculin A treated HEK-293 cells were subjected to SDS PAGE followed by western blot with 80462-1-RR (Phospho-AKT1 (Ser473) antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with GAPDH antibody as loading control.

• 

  FC experiment of NIH/3T3 using 80462-1-RR

  1X10^6 NIH/3T3 cells untreated (dashed line) or treated with Calyculin A (red) were intracellularly stained with 0.5 ug Anti-Human Phospho-AKT1 (Ser473) (80462-1-RR, Clone:2M10) and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) at dilution 1:1000, or 0.5 ug Control Antibody (blue). Cells were fixed with 4% PFA and permeabilized with 90% MeOH.

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Tested Applications

• Positive WB detected in: HeLa cells, HEK-293T cells, HEK-293 cells, NIH/3T3 cells, IGF-1 treated HEK-293T cells, Calyculin A treated HEK-293 cells, Calyculin A treated HeLa cells, Calyculin A treated NIH/3T3 cells
• Positive FC detected in: Calyculin A treated NIH/3T3 cells

Recommended dilution

• Western Blot (WB): WB : 1:2000-1:10000
• Flow Cytometry (FC): FC : 0.50 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 2 publications below

Product Information

80462-1-RR targets Phospho-AKT1 (Ser473) in WB, FC, ELISA applications and shows reactivity with Human, mouse samples.

• Tested Reactivity: Human, mouse
• Cited Reactivity: human, mouse
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: Peptide
• Full Name: v-akt murine thymoma viral oncogene homolog 1
• Observed Molecular Weight: 56-62 kDa
• GenBank Accession Number: NM_005163
• Gene Symbol: AKT1
• Gene ID (NCBI): 207
• RRID: AB_2918894
• 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). 80462-1-RR specifically recognizes AKT1 phosphorylated at Ser473.

Publications

Species	Application	Title
human	WB	ACS Omega; Interpreting the Pharmacological Mechanisms of Sho-saiko-to on Thyroid Carcinoma through Combining Network Pharmacology and Experimental Evaluation.; Authors - Kun Wang
mouse	WB	Biol Pharm Bull; Dictamnine ameliorates DNFB-induced atopic dermatitis like skin lesions in mice by inhibiting M1 macrophage polarization and promoting autophagy; Authors - Yihan Huang