Phospho-AKT (Ser473) Recombinant antibody

Phospho-AKT (Ser473) Uni-rAb^TM Recombinant Antibody for WB, FC (Intra), ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human and More (5)

Applications

WB, IHC, IF, FC (Intra), ELISA

Conjugate

Unconjugated

Publications

101

CloneNo.

2E17

Cat no : 80455-1-RR

Synonyms

AKT (Ser473), p AKT, p AKT (Ser473), p-AKT, p-AKT (Ser473)

---

Validation Data Gallery

• 

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

---

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 (Intra) detected in: Calyculin A treated HEK-293 cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 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 98 publications below
• IHC: See 8 publications below
• IF: See 2 publications below

Product Information

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

• Tested Reactivity: human
• Cited Reactivity: human, mouse, rat, pig, rabbit, 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	Cell Rep Med; Reprograming immunosuppressive microenvironment by eIF4G1 targeting to eradicate pancreatic ductal adenocarcinoma; Authors - Lihong He
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
rat	WB	Cell Mol Life Sci; BOP1 contributes to the activation of autophagy in polycystic ovary syndrome via nucleolar stress response; Authors - Rui Ji
human,mouse	WB,IHC	Phytomedicine; Tiliroside disrupted iron homeostasis and induced ferroptosis via directly targeting calpain-2 in pancreatic cancer cells; Authors - Maolei Xu
mouse	WB	Phytomedicine; Mechanism of Prunella vulgaris L. and luteolin in restoring Tfh/Tfr balance and alleviating oxidative stress in Graves' disease; Authors - Yunnan Zhang