Recombinant Anti-AKT1 + AKT2 + AKT3 (phospho S472 + S473 + S474) antibody [EPR18853] (ab192623)
![Western blot - Anti-AKT1 + AKT2 + AKT3 (phospho S472 + S473 + S474) antibody [EPR18853] (ab192623)](https://www.abcam.com/ps/products/192/ab192623/Images/ab192623-330826-anti-akt3-phospho-s472-akt2-phospho-s474-akt1-phospho-s473-antibody-epr18853-western-blot.jpg "Western blot - Anti-AKT1 + AKT2 + AKT3 (phospho S472 + S473 + S474) antibody [EPR18853] (ab192623)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18853] to AKT1 + AKT2 + AKT3 (phospho S472 + S473 + S474)
- Suitable for: ICC/IF, WB, IP, Dot blot
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-AKT1 + AKT2 + AKT3 (phospho S472 + S473 + S474) antibody [EPR18853]
See all AKT1 + AKT2 + AKT3 primary antibodies -
Description
Rabbit monoclonal [EPR18853] to AKT1 + AKT2 + AKT3 (phospho S472 + S473 + S474) -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, WB, IP, Dot blotmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: MCF7 whole cell lysate treated with 100ng/ml IGF-1 for 15 minutes; PC-12 and NIH/3T3 whole cell lysates treated with 100ng/ml PDGF for 60 minutes. ICC/IF: NIH/3T3 cells treated with PDGF (100 ng/ml) for 1 hour. IP: NIH/3T3 treated with 50ng/ml PDGF for 40min whole cell lysate.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR18853 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab192623 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| ICC/IF |
1/100.
|
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| WB |
1/1000. Detects a band of approximately 56 kDa (predicted molecular weight: 56 kDa).
|
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| IP |
1/40.
|
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| Dot blot |
1/1000.
|
| Notes |
|---|
|
ICC/IF
1/100. |
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WB
1/1000. Detects a band of approximately 56 kDa (predicted molecular weight: 56 kDa). |
|
IP
1/40. |
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Dot blot
1/1000. |
Target
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Function
IGF-1 leads to the activation of AKT3, which may play a role in regulating cell survival. Capable of phosphorylating several known proteins. Truncated isoform 2/PKB gamma 1 without the second serine phosphorylation site could still be stimulated but to a lesser extent. -
Tissue specificity
In adult tissues, it is highly expressed in brain, lung and kidney, but weakly in heart, testis and liver. In fetal tissues, it is highly expressed in heart, liver and brain and not at all in kidney. -
Sequence similarities
Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. RAC subfamily.
Contains 1 AGC-kinase C-terminal domain.
Contains 1 PH domain.
Contains 1 protein kinase domain. -
Domain
Binding of the PH domain to the phosphatidylinositol 3-kinase alpha (PI(3)K) results in its targeting to the plasma membrane. -
Post-translational
modificationsPhosphorylation on Thr-305 and Ser-472 is required for full activity (By similarity). Phosphorylated upon DNA damage, probably by ATM or ATR.
Ubiquitinated. When fully phosphorylated and translocated into the nucleus, undergoes 'Lys-48'-polyubiquitination catalyzed by TTC3, leading to its degradation by the proteasome. -
Cellular localization
Cytoplasm. Membrane. Membrane-associated after cell stimulation leading to its translocation. - Information by UniProt
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Database links
- Entrez Gene: 10000 Human
- Entrez Gene: 207 Human
- Entrez Gene: 208 Human
- Entrez Gene: 11651 Mouse
- Entrez Gene: 11652 Mouse
- Entrez Gene: 23797 Mouse
- Entrez Gene: 24185 Rat
- Entrez Gene: 25233 Rat
see all -
Alternative names
- AKT antibody
- AKT1 antibody
- AKT1 kinase antibody
see all
Images
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Western blot - Anti-AKT1 + AKT2 + AKT3 (phospho S472 + S473 + S474) antibody [EPR18853] (ab192623)All lanes : Anti-AKT1 + AKT2 + AKT3 (phospho S472 + S473 + S474) antibody [EPR18853] (ab192623) at 1/1000 dilution
Lane 1 : LNCaP (Human prostate carcinoma epithelial cell) whole cell lysates
Lane 2 : LNCaP (Human prostate carcinoma epithelial cell) treated with 0.1 uM Calyculin A for 30 minutes whole cell lysates
Lane 3 : A549 (Human lung carcinoma epithelial cell) whole cell lysates
Lane 4 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysates
Lane 5 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates
Lane 6 : HUVEC (Human umbilical vein endothelial cell) whole cell lysates
Lane 7 : C2C12 (Mouse myoblasts myoblast) whole cell lysates
Lane 8 : Mouse brain lysates
Lane 9 : Rat heart lysates
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 56 kDa
Observed band size: 56 kDa
Exposure time: 50 secondsThe basal expression level of AKT1 (phospho S473) varies in different cell lines reported by PMID: 19372546. But to detect clear signal, treatment is strongly recommended when using this antibody.
Blocking and Diluting buffer: 5% NFDM/TBST
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![Immunocytochemistry/ Immunofluorescence - Anti-AKT1 + AKT2 + AKT3 (phospho S472 + S473 + S474) antibody [EPR18853] (ab192623)](https://www.abcam.com/ps/products/192/ab192623/Images/ab192623-267934-anti-akt3-phospho-s472-akt2-phospho-s474-akt1-phospho-s473-antibody-epr18853-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-AKT1 + AKT2 + AKT3 (phospho S472 + S473 + S474) antibody [EPR18853] (ab192623)Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells, untreated or treated with PDGF (100 ng/ml) for 1 hour, labeling AKT3 (phospho S472) + AKT2 (phospho S474) + AKT1 (phospho S473) with ab192623 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
The signal increased after treatment with PDGF (100 ng/ml) for 1 hour on NIH/3T3 cells.
The nuclear counter stain is DAPI (blue).
Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
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![Dot Blot - Anti-AKT1 + AKT2 + AKT3 (phospho S472 + S473 + S474) antibody [EPR18853] (ab192623)](https://www.abcam.com/ps/products/192/ab192623/Images/ab192623-267937-anti-akt3-phospho-s472-akt2-phospho-s474-akt1-phospho-s473-antibody-epr18853-dot-blot.jpg)
Dot Blot - Anti-AKT1 + AKT2 + AKT3 (phospho S472 + S473 + S474) antibody [EPR18853] (ab192623)Dot blot analysis of AKT3 (phospho S472) labeled with ab192623 at 1/1000 dilution.
Lane 1: AKT3 (phospho S472) phospho peptide;
Lane 2: AKT3 non-phospho peptide;
Lane 3: AKT1 (phospho S473) phospho peptide;
Lane 4: AKT2 (phospho S474) phospho peptide.
Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated (ab97051) at 1/100000 dilution was used as secondary antibody.
Blocking and diluting buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
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![Western blot - Anti-AKT1 + AKT2 + AKT3 (phospho S472 + S473 + S474) antibody [EPR18853] (ab192623)](https://www.abcam.com/ps/products/192/ab192623/Images/ab192623-267936-anti-akt3-phospho-s472-akt2-phospho-s474-akt1-phospho-s473-antibody-epr18853-western-blot.jpg)
Western blot - Anti-AKT1 + AKT2 + AKT3 (phospho S472 + S473 + S474) antibody [EPR18853] (ab192623)All lanes : Anti-AKT1 + AKT2 + AKT3 (phospho S472 + S473 + S474) antibody [EPR18853] (ab192623) at 1/1000 dilution
Lane 1 : Untreated MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
Lane 2 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate treated with 100ng/ml IGF-1 for 15 minutes
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 56 kDa
Observed band size: 56 kDa
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
Phosphorylation of AKT at S473 can be induced by IGF-1 treatment according to the literature (PMID: 23638184).
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![Western blot - Anti-AKT1 + AKT2 + AKT3 (phospho S472 + S473 + S474) antibody [EPR18853] (ab192623)](https://www.abcam.com/ps/products/192/ab192623/Images/ab192623-267935-anti-akt3-phospho-s472-akt2-phospho-s474-akt1-phospho-s473-antibody-epr18853-western-blot.jpg)
Western blot - Anti-AKT1 + AKT2 + AKT3 (phospho S472 + S473 + S474) antibody [EPR18853] (ab192623)All lanes : Anti-AKT1 + AKT2 + AKT3 (phospho S472 + S473 + S474) antibody [EPR18853] (ab192623) at 1/1000 dilution
Lane 1 : Untreated PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate treated with 100ng/ml PDGF for 60 minutes
Lane 3 : Untreated NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
Lane 4 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate treated with 100ng/ml PDGF for 60 minutes
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 56 kDa
Observed band size: 56 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1 and 2: 15 seconds; Lane 3 and 4: 3 minutes.
Phosphorylation of AKT can be induced by PDGF treatment according to the literature (PMID: 10984605 and 7774014).
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![Immunoprecipitation - Anti-AKT1 + AKT2 + AKT3 (phospho S472 + S473 + S474) antibody [EPR18853] (ab192623)](https://www.abcam.com/ps/products/192/ab192623/Images/ab192623-267933-anti-akt3-phospho-s472-akt2-phospho-s474-akt1-phospho-s473-antibody-epr18853-immunoprecipitation.jpg)
Immunoprecipitation - Anti-AKT1 + AKT2 + AKT3 (phospho S472 + S473 + S474) antibody [EPR18853] (ab192623)AKT3 (phospho S472) was immunoprecipitated from 0.35 mg of NIH/3T3 (Mouse embryonic fibroblast cell line) treated with 50ng/ml PDGF for 40min whole cell lysate with ab192623 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab192623 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.
Lane 1: NIH/3T3 treated with 50ng/ml PDGF for 40min whole cell lysate, 10μg (Input).
Lane 2: ab192623 IP in NIH/3T3 treated with 50ng/ml PDGF for 40min whole cell lysate.
Lane 3: Rabbit IgG,monoclonal [EPR25A]-Isotype Control (ab172730) instead of ab192623 in NIH/3T3 treated with 50ng/ml PDGF for 40min whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 5 seconds.
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![Anti-AKT1 + AKT2 + AKT3 (phospho S472 + S473 + S474) antibody [EPR18853] (ab192623)](https://www.abcam.com/ps/products/192/ab192623/Images/ab192623-7-benefits-of-recombinant-antibodies.png)
Anti-AKT1 + AKT2 + AKT3 (phospho S472 + S473 + S474) antibody [EPR18853] (ab192623)
Protocols
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (26)
ab192623 has been referenced in 26 publications.
- Liu J et al. Protective effect of puerarin against burn-induced heart injury in rats. Exp Ther Med 20:275-282 (2020). PubMed: 32536996
- Zhu X et al. Ophiopogonin D suppresses TGF-ß1-mediated metastatic behavior of MDA-MB-231 breast carcinoma cells via regulating ITGB1/FAK/Src/AKT/ß-catenin/MMP-9 signaling axis. Toxicol In Vitro 69:104973 (2020). PubMed: 32818624
- Li D et al. Circular RNA CDR1as Exerts Oncogenic Properties Partially through Regulating MicroRNA 641 in Cholangiocarcinoma. Mol Cell Biol 40:N/A (2020). PubMed: 32423991
- Xiao F et al. Tobacco extracts promote PD-L1 expression and enhance malignant biological differences via mTOR in gefitinib-resistant cell lines. Thorac Cancer 11:2237-2251 (2020). PubMed: 32558328
- Wu HK et al. PHLPP2 is regulated by competing endogenous RNA network in pathogenesis of colon cancer. Aging (Albany NY) 12:12812-12840 (2020). PubMed: 32633726
