Anti-AKT2 (phospho S474) antibody (ab38513)
- Datasheet
- Specific References (11)
- Protocols
Overview
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Product nameAnti-AKT2 (phospho S474) antibody
See all AKT2 primary antibodies -
DescriptionRabbit polyclonal to AKT2 (phospho S474)
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Host speciesRabbit
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SpecificityAkt2 (phospho-Ser474) antibody detects endogenous levels of Akt2 only when phosphorylated at serine 474. The region of AKT2 surrounding S474 has a high degree of similarity to the corresponding regions in AKT1 and AKT3 and thus may cross react with these proteins if phosphorylated on the corresponding serine residue.
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Tested applicationsSuitable for: WB, IHC-P, ICC/IFmore details
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Species reactivityReacts with: Mouse, Rat, Human
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Immunogen
Synthetic peptide within Human AKT2 aa 432-481 (C terminal) (phospho S474). The exact sequence is proprietary. The antiserum was produced against synthesized phosphopeptide derived from human Akt2 around the phosphorylation site of serine 474.
Sequence:SEVDTRYFD DEFTAQSITI TPPDRYDSLG LLELDQRTHF PQFSYSASIR E
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Positive control
- Lung carcinoma tissue slides, A2780 cell extract and EGF treated 293 cell lysate.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
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Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Constituents: 50% Glycerol, 0.87% Sodium chloride, 49% PBS
PBS without Mg2+ and Ca2+. -
Concentration information loading... -
PurityProtein A purified
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Purification notesThe antibody was affinity purified from rabbit antiserum by affinity chromatography using epitope-specific phosphopeptide. The antibody against non-phosphopeptide was removed by chromatography using non-phosphopeptide corresponding to the phosphorylation site.
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Compatible Secondaries
Applications
Our Abpromise guarantee covers the use of ab38513 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 |
|---|---|---|
| WB | 1/500 - 1/1000. Predicted molecular weight: 55 kDa. | |
| IHC-P | 1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. | |
| ICC/IF | Use at an assay dependent concentration. |
Target
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FunctionGeneral protein kinase capable of phosphorylating several known proteins.
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Tissue specificityExpressed in all human cell types so far analyzed.
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Sequence similaritiesBelongs 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. -
Post-translational
modificationsPhosphorylation on Thr-309 and Ser-474 is required for full activity.
Ubiquitinated; undergoes both 'Lys-48'- and 'Lys-63'-linked polyubiquitination. TRAF6-induced 'Lys-63'-linked AKT2 ubiquitination. When fully phosphorylated and translocated into the nucleus, undergoes 'Lys-48'-polyubiquitination catalyzed by TTC3, leading to its degradation by the proteasome. - Information by UniProt
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Database links
- Entrez Gene: 208 Human
- Entrez Gene: 11652 Mouse
- Entrez Gene: 25233 Rat
- Omim: 164731 Human
- SwissProt: P31751 Human
- SwissProt: Q60823 Mouse
- SwissProt: P47197 Rat
- Unigene: 631535 Human
see all -
Alternative names
- Akt2 antibody
- AKT2_HUMAN antibody
- HIHGHH antibody
see all
Images
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Western blot - Anti-AKT2 (phospho S474) antibody (ab38513)All lanes : Anti-AKT2 (phospho S474) antibody (ab38513)
Lane 1 : 293 cells treated for 30 min with 200ng/ml EGF
Lane 2 : 293 cells treated for 30 min with 200ng/ml EGF with Phospho peptide
Predicted band size: 55 kDa -
Immunocytochemistry/ Immunofluorescence - Anti-AKT2 (phospho S474) antibody (ab38513)This image is courtesy of an anonymous Abreviewab38513 staining AKT2 (phospho S474) in 293T cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with 0.1% Tween and blocked with 3% BSA for 1 hour at room temperature. Samples were incubated with primary antibody (1/20 in PBS + 3% BSA) for 2 hours. An Alexa Fluor® 555-conjugated donkey anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody.
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Western blot - Anti-AKT2 (phospho S474) antibody (ab38513)All lanes : Anti-AKT2 (phospho S474) antibody (ab38513) at 1/500 dilution
Lane 1 : Extract of A2780 cells.
Lane 2 : Extract of A2780 cells + TNF alpha.
Lane 3 : Extract of A2780 cells + UV.
Lysates/proteins at 30 µg per lane.
Secondary
All lanes : Alkaline Phosphatase AffiniPure Goat Anti-Rabbit IgG (H+L)
Predicted band size: 55 kDa
Observed band size: 60 kDa (why is the actual band size different from the predicted?)
Lanes can be loaded with 5-30µg of total protein. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT2 (phospho S474) antibody (ab38513)This image shows human lung carcinoma tissue stained with ab38513 at 1/50 dilution.
Image on left: untreated.
Image on right: treated with phosphopeptide at 1µg/ml (negative control).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT2 (phospho S474) antibody (ab38513)This image is courtesy of an anonymous Abreviewab38513 staining AKT2 in human infantile fibromatosis tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% FBS/BSA for 3 hours at room temperature; antigen retrieval was by heat mediation in Tris pH 9. Samples were incubated with primary antibody (1/150 in TBS + 1% BSA + 1% FBS) for 16 hours. An undiluted HRP-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody.
Protocols
References
This product has been referenced in:
- Chen Y et al. Testicular orphan receptor 4 promotes tumor progression and implies poor survival through AKT3 regulation in seminoma. Cancer Sci 109:384-394 (2018). Read more (PubMed: 29197138) »
- Yang Y et al. Regulation of macrophage migration in ischemic mouse hearts via an AKT2/NBA1/SPK1 pathway. Oncotarget 8:115345-115359 (2017). Read more (PubMed: 29383164) »
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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"