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    akt2-phospho-s474-antibody-ab38513.pdf

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Signal Transduction Protein Phosphorylation Ser / Thr Kinases PKB / AKT
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Anti-AKT2 (phospho S474) antibody (ab38513)

  • Datasheet
Reviews (6) Submit a question References (23)

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Western blot - Anti-AKT2 (phospho S474) antibody (ab38513)
  • Western blot - Anti-AKT2 (phospho S474) antibody (ab38513)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT2 (phospho S474) antibody (ab38513)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT2 (phospho S474) antibody (ab38513)

Key features and details

  • Rabbit polyclonal to AKT2 (phospho S474)
  • Suitable for: WB, IHC-P
  • Reacts with: Human
  • Isotype: IgG

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Overview

  • Product name

    Anti-AKT2 (phospho S474) antibody
    See all AKT2 primary antibodies
  • Description

    Rabbit polyclonal to AKT2 (phospho S474)
  • Host species

    Rabbit
  • Specificity

    Akt2 (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 applications

    Suitable for: WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
  • 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

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: 50% Glycerol, 0.87% Sodium chloride, 49% PBS

    PBS without Mg2+ and Ca2+.
  • Concentration information loading...
  • Purity

    Protein A purified
  • Purification notes

    The 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.
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Kinases
    • PKB / AKT
    • Cancer
    • Signal transduction
    • Protein phosphorylation
    • Serine/threonine kinases
    • AKT
    • Cardiovascular
    • Atherosclerosis
    • Diabetes associated
    • Cardiovascular
    • Heart
    • Cardiac metabolism
    • Metabolism
    • Types of disease
    • Diabetes
    • Metabolism
    • Types of disease
    • Cancer
    • Metabolism
    • Types of disease
    • Heart disease

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat Anti-Rabbit IgG H&L (Alkaline Phosphatase) (ab6722)
  • Immunizing Peptide (Blocking)

    • Recombinant human AKT2 protein (ab79798)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant human AKT2 protein (ab79798)

Applications

The Abpromise guarantee

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 (2)
1/500 - 1/1000. Predicted molecular weight: 55 kDa.Can be blocked with Recombinant human AKT2 protein (ab79798).
IHC-P (2)
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Notes
WB
1/500 - 1/1000. Predicted molecular weight: 55 kDa.Can be blocked with Recombinant human AKT2 protein (ab79798).
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Target

  • Function

    General protein kinase capable of phosphorylating several known proteins.
  • Tissue specificity

    Expressed in all human cell types so far analyzed.
  • 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.
  • Post-translational
    modifications

    Phosphorylation 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.
  • Target information above from: UniProt accession P31751 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 208 Human
    • Omim: 164731 Human
    • SwissProt: P31751 Human
    • Unigene: 631535 Human
    • Alternative names

      • Akt2 antibody
      • AKT2_HUMAN antibody
      • HIHGHH antibody
      • Murine thymoma viral (v-akt) homolog 2 antibody
      • murine thymoma viral (v-akt) homolog-2 antibody
      • Oncogene AKT2 protein kinase B beta antibody
      • PKB antibody
      • PKB beta antibody
      • PKBB antibody
      • PKBBETA antibody
      • PRKBB antibody
      • Protein kinase Akt 2 antibody
      • Protein kinase Akt-2 antibody
      • Protein kinase B beta antibody
      • RAC beta antibody
      • rac protein kinase beta antibody
      • RAC-BETA antibody
      • RAC-beta serine/threonine-protein kinase antibody
      • RAC-PK-beta antibody
      • RACbeta antibody
      • v akt murine thymoma viral oncogene homolog 2 antibody
      • V-AKT murine thymoma viral oncogene homolog 2 antibody
      see all

    Images

    • Western blot - Anti-AKT2 (phospho S474) antibody (ab38513)
      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

    • Western blot - Anti-AKT2 (phospho S474) antibody (ab38513)
      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)
      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).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT2 (phospho S474) antibody (ab38513)
      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.

      See Abreview

    Protocols

    • Western blot protocols
    • Immunohistochemistry protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    References (23)

    Publishing research using ab38513? Please let us know so that we can cite the reference in this datasheet.

    ab38513 has been referenced in 23 publications.

    • Wang Y  et al. Inhibition of bone morphogenetic protein receptor 2 suppresses pancreatic ductal adenocarcinoma growth by regulating GRB2/PI3K/AKT axis. Ann Transl Med 9:557 (2021). PubMed: 33987255
    • Hayashi-Hori M  et al. Therapeutic Effect of Rapamycin on Aortic Dissection in Mice. Int J Mol Sci 21:N/A (2020). PubMed: 32397282
    • Zhao J  et al. Metoprolol alleviates arginine vasopressin-induced cardiomyocyte hypertrophy by upregulating the AKT1-SERCA2 cascade in H9C2 cells. Cell Biosci 10:72 (2020). PubMed: 32489586
    • Duan L  et al. Magnetic Targeting of HU-MSCs in the Treatment of Glucocorticoid-Associated Osteonecrosis of the Femoral Head Through Akt/Bcl2/Bad/Caspase-3 Pathway. Int J Nanomedicine 15:3605-3620 (2020). PubMed: 32547017
    • Xu X  et al. Triptolide inhibits angiogenesis in microvascular endothelial cells through regulation of miR-92a. J Physiol Biochem 75:573-583 (2019). PubMed: 31691162
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-2 of 2 Abreviews or Q&A

    Western blot abreview for Anti-AKT2 (phospho S474) antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    25 µg
    Gel Running Conditions
    Reduced Denaturing (12)
    Sample
    Pig Cell lysate - whole cell (PAE cells)
    Specification
    PAE cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Feb 25 2014

    Western blot abreview for Anti-AKT2 (phospho S474) antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Mouse mammary cell)
    Loading amount
    100 µg
    Specification
    Mouse mammary cell
    Treatment
    5ng/ml TGF beta for different time periods
    Gel Running Conditions
    Reduced Denaturing (10.0% gel)
    Blocking step
    BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Aug 31 2011

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