APPLICATION
Application Note
*Optimal dilutions/concentrations should be determined by the researcher.
| Application |
Recommended Dilution |
| 1:500-1:3000 |
| 1:100-1:1000 |
| Assay dependent |
| 1:100-1:500 |
Not tested in other applications.
Calculated MW
Positive Control
MCF-7(500 nM PMA treatment for 1 hr) , MCF-7(500 nM PMA treatment for 2 hr) , MCF-7(500 nM PMA treatment for 4 hr) , MCF-7(500 nM PMA treatment for 6 hr) , HUVEC (5 ng/ml VEGF treatment for 10 min) , PC-12 , Rat2
Product Note
This antibody is raised against human AKT1 phosphorylated at Ser473. Based on sequence homology, it is predicted to react with AKT2 and AKT3 when phosphorylated at the corresponding residues - AKT2 Ser474 and AKT3 Ser472.
Predict Reactivity
Zebrafish, Sheep, Bovine, Cat, Chicken, Rhesus Monkey(>80% identity)
PROPERTIES
Form
Liquid
Buffer
PBS, 1% BSA, 20% Glycerol
Preservative
0.025% ProClin 300
Storage
Store as concentrated solution. Centrifuge briefly prior to opening vial. For short-term storage (1-2 weeks), store at 4ºC. For long-term storage, aliquot and store at -20ºC or below. Avoid multiple freeze-thaw cycles.
Concentration
1 mg/ml (Please refer to the vial label for the specific concentration.)
Antigen Species
Human
Immunogen
Carrier-protein conjugated synthetic peptide surrounding phospho Ser473 of human AKT. The exact sequence is proprietary.
Purification
Purified by antigen-affinity chromatography.
Conjugation
Unconjugated
RRID
AB_2885773
Note
For laboratory research use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. Not for animal or human consumption.
Purchasers shall not, and agree not to enable third parties to, analyze, copy, reverse engineer or otherwise attempt to determine the structure or sequence of the product.
TARGET
Synonyms
AKT serine/threonine kinase 1 , AKT , CWS6 , PKB , PKB-ALPHA , PRKBA , RAC , RAC-ALPHA
Cellular Localization
Cytoplasm , Nucleus , Cell membrane
Background
The serine-threonine protein kinase encoded by the AKT1 gene is catalytically inactive in serum-starved primary and immortalized fibroblasts. AKT1 and the related AKT2 are activated by platelet-derived growth factor. The activation is rapid and specific, and it is abrogated by mutations in the pleckstrin homology domain of AKT1. It was shown that the activation occurs through phosphatidylinositol 3-kinase. In the developing nervous system AKT is a critical mediator of growth factor-induced neuronal survival. Survival factors can suppress apoptosis in a transcription-independent manner by activating the serine/threonine kinase AKT1, which then phosphorylates and inactivates components of the apoptotic machinery. Multiple alternatively spliced transcript variants have been found for this gene. [provided by RefSeq]
Database
Research Area
DATA IMAGES
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GTX128414 ICC/IF Image
AKT (phospho Ser473) antibody detects AKT (phospho Ser473) protein by immunofluorescent analysis.Sample: NIH3T3 cells were fixed in 4% paraformaldehyde at RT for 15 min.Green: AKT (phospho Ser473) stained by AKT (phospho Ser473) antibody (GTX128414) diluted at 1:500.Blue: Hoechst 33342 staining.Scale bar= 10 μm.
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GTX128414 WB Image
MCF-7 cells were untreated or treated with 500 nM PMA for 1, 2, 4 and 6 hrs. Various whole cell extracts (30 μg) were separated by 7.5% SDS-PAGE, and the membrane was blotted with AKT (phospho Ser473) antibody (GTX128414) diluted at 1:500. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.
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GTX128414 IP Image
Immunoprecipitation of Akt1/2/3 (phospho Ser473/474/472) protein from 293T whole cell extracts using 5 μg of Akt1/2/3 (phospho Ser473/474/472) antibody (GTX128414).
Western blot analysis was performed using Akt1/2/3 (phospho Ser473/474/472) antibody (GTX128414).
EasyBlot anti-Rabbit IgG (GTX221666-01) was used as a secondary reagent.
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GTX128414 WB Image
Untreated (–) and treated (+) NIH-3T3 whole cell extracts (30 μg) were separated by 10% SDS-PAGE, and the membrane was blotted with AKT (phospho Ser473) antibody (GTX128414) diluted at 1:500. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.
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GTX128414 WB Image
Untreated (–) and treated (+) HUVEC whole cell extracts (30 μg) were separated by 7.5% SDS-PAGE, and the membrane was blotted with AKT (phospho Ser473) antibody (GTX128414) diluted at 1:500. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.
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GTX128414 WB Image
Various whole cell extracts (30 μg) were separated by 10% SDS-PAGE, and the membrane was blotted with AKT (phospho Ser473) antibody (GTX128414) diluted at 1:500. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody, and the signal was developed with Trident ECL plus-Enhanced.
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GTX128414 WB Image
Untreated (–) and treated (+) MCF-7 whole cell extracts (30 μg) were separated by 10% SDS-PAGE, and the membrane was blotted with AKT (phospho Ser473) antibody (GTX128414) diluted at 1:500. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.
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GTX128414 WB Image
Untreated (–) and treated (+) MCF-7 whole cell extracts (30 μg) were separated by 10% SDS-PAGE, and the membrane was blotted with AKT (phospho Ser473) antibody (GTX128414) diluted at 1:500. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.
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GTX128414 WB Image
The data was published in the 2021 in Cancer Drug Resist. PMID: 35582384
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GTX128414 WB Image
The data was published in the journal Antioxidants (Basel) in 2019.PMID: 31906147
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GTX128414 WB Image
The data was published in the journal Int J Mol Sci in 2017. PMID: 28837077
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GTX128414 WB Image
The data was published in the journal Biomed Res Int in 2015. PMID: 26557697
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GTX128414 WB Image
The data was published in the journal Cell Death Dis in 2014. PMID: 24457964
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REFERENCE
REVIEW
AKT (phospho Ser473) antibody
Cat. No. GTX128414
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Rating
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( Average 4 based on 3 users reviews)
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Application
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Western Blot(WB)
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( Average 4 based on 3 users reviews)
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Date :
Anonymous submitted on 05-Feb-2024
Application Tested :
WB
Sample Species :
Hu
Sample :
RWPE-1 cells with/without treatment
Amount used :
20μg
Blocking :
Blocking solution, 4°C, 0.1Hr
Primary Antibody :
1:3000, 4°C, 17Hr
Date :
Anonymous submitted on 16-Dec-2020
Application Tested :
WB
Sample Species :
Hu
Sample :
SW1353 cells
Amount used :
10 μg
Blocking :
Block Ace (DS Parma Biomedical Co. Ltd., Osaka, Japan), 25°C, 1Hr
Primary Antibody :
1:1000, 4°C, 16Hr
Date :
Anonymous submitted on 14-Apr-2016
Application Tested :
WB
Sample Species :
Hu
Sample :
HUVEC cells
Amount used :
30μg
Blocking :
5% milk, 25°C, 1Hr
Primary Antibody :
1:1500, 4°C, 16Hr
