Phospho-AKT (Ser473) Polyclonal antibody

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  WB analysis using 28731-1-AP

  Non-treated PC-3 and Calyculin A treated PC-3 cells were subjected to SDS PAGE followed by western blot with 28731-1-AP (Phospho-AKT (Ser473) antibody) at dilution of 1:3000 incubated at 4℃ overnight. The membrane was stripped and re-blotted with GAPDH antibody as loading control.

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  WB analysis using 28731-1-AP

  Non-treated NIH/3T3 and PDGF (HZ-1215) treated NIH/3T3 cells were subjected to SDS PAGE followed by western blot with 28731-1-AP (Phospho-AKT (Ser473) antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

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  IF Staining of HeLa using 28731-1-AP

  Immunofluorescent analysis of (4% PFA) fixed Calyculin A treated HeLa cells using Phospho-AKT (Ser473) antibody (28731-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2).

Phospho-AKT (Ser473) Polyclonal Antibody for WB, IF/ICC, ELISA

Cat No. 28731-1-AP

Publications(198)

Reviews (2)

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat and More (3)

Applications

WB, IHC, IF/ICC, ELISA

Synonyms

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

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Tested Applications

• Positive WB detected in: Calyculin A treated PC-3 cells, PDGF treated NIH/3T3 cells
• Positive IF/ICC detected in: Calyculin A treated HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:1000-1:5000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:50-1:500

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 196 publications below
• IHC: See 7 publications below
• IF: See 4 publications below

Product Information

28731-1-AP targets Phospho-AKT (Ser473) in WB, IHC, IF/ICC, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse, rat, pig, rabbit, sheep
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• 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_2881201
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• UNIPROT ID: P31749
• 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

Phosphorylation on Thr-308, Ser-473 and Tyr-474 is required for full activity. Activated TNK2 phosphorylates it on Tyr-176 resulting in its binding to the anionic plasma membrane phospholipid PA (PubMed:20333297). This phosphorylated form localizes to the cell membrane, where it is targeted by PDPK1 and PDPK2 for further phosphorylations on Thr-308 and Ser-473 leading to its activation (PubMed:9512493). Ser-473 phosphorylation by mTORC2 favors Thr-308 phosphorylation by PDPK1 (PubMed:21464307, PubMed:8978681). AKT1 Thr-308 and Ser-473 is Phosphorylated by IKBKE and TBK1 (PubMed:15718470, PubMed:18456494, PubMed:20481595, PubMed:8978681). Ser-473 phosphorylation is enhanced by interaction with AGAP2 isoform 2 (PIKE-A) (PubMed:14761976). Ser-473 phosphorylation is enhanced in focal cortical dysplasias with Taylor-type balloon cells (PubMed:17013611). Ser-473 phosphorylation is enhanced by signaling through activated FLT3 (By similarity). Ser-473 is dephosphorylated by PHLPP (PubMed:28147277) and Ser-473 by PP2A phosphatase (PubMed:21329884).

Publications

• Human
• Mouse
• Rat
• Monkey
• Pig

Species	Application	Title
mouse	WB	Toxicology; Toxicity mechanism of peri-implantation pesticide beta-cypermethrin exposure on endometrial remodeling in early pregnant mice; Authors - Yong-Jiang Zhou
mouse	WB	J Neuroinflammation; TREM2 affects DAM-like cell transformation in the acute phase of TBI in mice by regulating microglial glycolysis; Authors - Lin Wang
mouse	WB	Phytomedicine; Combinational application of the natural products 1-deoxynojirimycin and morin ameliorates insulin resistance and lipid accumulation in prediabetic mice; Authors - Xinxiu Ren
human	WB	Biomed Pharmacother; Preclinical characterization of danatinib as a novel FLT3 inhibitor with excellent efficacy against resistant acute myeloid leukemia; Authors - Shan-Liang Sun
human	WB	Cell Biosci; Serine protease PRSS56, a novel cancer-testis antigen activated by DNA hypomethylation, promotes colorectal and gastric cancer progression via PI3K/AKT axis; Authors - Dandan Li
rat	WB	Biomed Pharmacother; Cinnamaldehyde protects donor heart from cold ischemia-reperfusion injury via the PI3K/AKT/mTOR pathway; Authors - Hongwen Lan

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive for providing their feedback.

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FH

Julia (Verified Customer) (07-24-2025)

Consistent results

• Applications: Western Blot
• Primary Antibody Dilution: 1:250

FH

Itziar (Verified Customer) (06-27-2022)

This antibody works very well with even little protein quantities

• Applications: Western Blot
• Primary Antibody Dilution: %5 BSA in TBST
• Cell Tissue Type: human pancreatic beta cells and Jurkat