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.
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
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.
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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.
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.
v akt murine thymoma viral oncogene homolog 2 antibody
Anti-AKT2 (phospho S474) antibody images
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 Abreview
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.
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).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT2 (phospho S474) antibody (ab38513)This image is courtesy of an anonymous Abreview
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.
References for Anti-AKT2 (phospho S474) antibody (ab38513)
This product has been referenced in:
Zhao HJ et al. MiR-194 Deregulation Contributes To Colorectal Carcinogenesis via Targeting AKT2 Pathway. Theranostics4:1193-208 (2014).
Read more (PubMed: 25285168) »
Glienke W et al. The dual PI3K/mTOR inhibitor NVP-BGT226 induces cell cycle arrest and regulates Survivin gene expression in human pancreatic cancer cell lines. Tumour Biol33:757-65 (2012).
Read more (PubMed: 22170433) »