APPLICATION
Application Note
*Optimal dilutions/concentrations should be determined by the researcher.
| Application |
Dilution |
| 1:500-1:3000 |
| 1:100-1:1000 |
| Assay dependent |
| 1:100-1:500 |
Not tested in other applications.
Calculated MW
Positive Control
PC-12 , Rat2 , MCF-7 , 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 , HUVEC (5 ng/ml VEGF treatment for 10 min) , NIH-3T3 , NIH-3T3 (starvation for 16 hr and 5 ng/ml PDGF treatment for 15 min)
Specificity/Sensitivity
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, Cat, Chicken, Xenopus laevis, Rhesus Monkey(>80% identity)
PROPERTIES
Form
Liquid
Buffer
1XPBS, 1% BSA, 20% Glycerol (pH7). 0.025% ProClin 300 was added as a preservative.
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
0.06 mg/ml (Please refer to the vial label for the specific concentration.)
Antigen Species
Human
Immunogen
Carrier-protein conjugated synthetic peptide corresponding to residues around human AKT (phospho Ser473). The exact sequence is proprietary.
Purification
Purified by antigen-affinity chromatography.
Conjugation
Unconjugated
Note
For laboratory use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. Not for animal or human consumption.
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 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
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 (+) 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
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 WB Image
Untreated (–) and treated (+) NIH-3T3 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
The WB analysis of AKT (phospho Ser473) antibody was published by Sun W and colleagues in the journal Cell Death Dis in 2014 .
PMID: 24457964
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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
The WB analysis of AKT (phospho Ser473) antibody was published by Vallejo-Flores G and colleagues in the journal Biomed Res Int in 2015.PMID: 26557697
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GTX128414 WB Image
The WB analysis of AKT (phospho Ser473) antibody was published by Lin JG and colleagues in the journal Int J Mol Sci in 2017.PMID: 28837077
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GTX128414 WB Image
Untreated (–) and treated (+) MCF-7 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, and the signal was developed with Trident ECL plus-Enhanced.
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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 1 users reviews)
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Application
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Western Blot(WB)
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( Average 4 based on 1 users reviews)
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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
