Product nameAnti-AKT2 (phospho S474) antibody
See all AKT2 primary antibodies
DescriptionRabbit polyclonal to AKT2 (phospho S474)
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.
Tested applicationsSuitable for: WB, IHC-P, ICC/IFmore details
Species reactivityReacts with: Mouse, Rat, Human
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.
SEVDTRYFD DEFTAQSITI TPPDRYDSLG LLELDQRTHF PQFSYSASIR E
- Lung carcinoma tissue slides, A2780 cell extract and EGF treated 293 cell lysate.
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.
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
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.
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.
|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.|
FunctionGeneral protein kinase capable of phosphorylating several known proteins.
Tissue specificityExpressed in all human cell types so far analyzed.
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.
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
- Akt2 antibody
- AKT2_HUMAN antibody
- HIHGHH antibody
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
ab38513 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.
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.
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.
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).
ab38513 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.
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) »