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  1. Link

    akt1-phospho-s473-antibody-ep2109y-ab81283.pdf

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Signal Transduction Protein Phosphorylation Ser / Thr Kinases PKB / AKT
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RecombinantRabMAb

Recombinant Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)

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Reviews (10)Q&A (1)References (481)

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Western blot - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)
  • Western blot - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)
  • Western blot - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)
  • Western blot - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)
  • Dot Blot - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)
  • Western blot - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)
  • Western blot - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)
  • Western blot - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)
  • Western blot - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)
  • In-Cell ELISA - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)
  • Western blot - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)
  • Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP2109Y] to AKT1 (phospho S473)
  • Suitable for: Dot blot, In-Cell ELISA, IHC-P, WB
  • Reacts with: Mouse, Rat, Human, Recombinant fragment

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Overview

  • Product name

    Anti-AKT1 (phospho S473) antibody [EP2109Y]
    See all AKT1 primary antibodies
  • Description

    Rabbit monoclonal [EP2109Y] to AKT1 (phospho S473)
  • Host species

    Rabbit
  • Specificity

    AKT1 (phospho S473) antibody (ab81283) detects AKT1 phosphorylated at Serine 473. The region of AKT1 surrounding S473 has a high degree of similarity to the corresponding regions in AKT2 and AKT3 and thus may cross react with these proteins if phosphorylated on the corresponding serine residue.

  • Tested applications

    Suitable for: Dot blot, In-Cell ELISA, IHC-P, WBmore details
    Unsuitable for: Flow Cyt,ICC/IF or IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human, Recombinant fragment
  • Immunogen

    Synthetic peptide corresponding to Human AKT1 (phospho S473).
    Database link: P31749
    (Peptide available as ab171724, ab217601)

  • Positive control

    • WB: HeLa (grown in serum free media overnight, then treated with 150nM Insulin for 5min) whole cell lysate; MCF7 (treated with CCCP) whole cell lysate; LNCaP (treated with 100nM Cacyculin A for 30min) whole cell lysate; NIH/3T3 (treated with PDGF) whole cell lysate; PC-12 whole cell lysate. IHC-P: Human cervical carcinoma tissue. Dot Blot: AKT1 (phospho S473) phospho peptide.
  • General notes

    Abcam recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor® 488 (ab150077).

    See other anti-rabbit secondary antibodies that can be used with this antibody.

    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
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

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: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EP2109Y
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Kinases
    • PKB / AKT
    • Epigenetics and Nuclear Signaling
    • Cell cycle
    • Apoptosis
    • Nuclear
    • Cancer
    • Signal transduction
    • Protein phosphorylation
    • Serine/threonine kinases
    • AKT
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Apoptosis
    • Cancer
    • Cell Death
    • Apoptosis
    • Metabolism

Associated products

  • Alternative Versions

    • HRP Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab194201)
    • Anti-AKT1 (phospho S473) antibody [EP2109Y] - BSA and Azide free (ab215873)
  • Assay kits

    • Akt Kinase Activity Assay Kit (ab139436)
    • AKT Activity Assay Kit (ab65786)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • HeLa Ser/Thr Phosphatase-Inhibited Whole Cell Lysate Set: Calyculin-Treated and Vehicle-Treated (ab135395)
  • Recombinant Protein

    • Recombinant human AKT1 protein (ab79792)
  • Related Products

    • Prestained Protein Ladder – Broad molecular weight (10-245 kDa) (ab116028)
    • DAPT, gamma-Secretase inhibitor (ab120633)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab81283 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
Dot blot
1/1000.
In-Cell ELISA
Use at an assay dependent concentration.
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB (5)
1/5000 - 1/10000. Predicted molecular weight: 56 kDa.

Abcam recommends using BSA as the blocking agent.

Notes
Dot blot
1/1000.
In-Cell ELISA
Use at an assay dependent concentration.
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB
1/5000 - 1/10000. Predicted molecular weight: 56 kDa.

Abcam recommends using BSA as the blocking agent.

Application notes
Is unsuitable for Flow Cyt,ICC/IF or IP.

Target

  • Function

    Plays a role as a key modulator of the AKT-mTOR signaling pathway controlling the tempo of the process of newborn neurons integration during adult neurogenesis, including correct neuron positioning, dendritic development and synapse formation (By similarity). General protein kinase capable of phosphorylating several known proteins. Phosphorylates TBC1D4. Signals downstream of phosphatidylinositol 3-kinase (PI(3)K) to mediate the effects of various growth factors such as platelet-derived growth factor (PDGF), epidermal growth factor (EGF), insulin and insulin-like growth factor I (IGF-I). Plays a role in glucose transport by mediating insulin-induced translocation of the GLUT4 glucose transporter to the cell surface. Mediates the antiapoptotic effects of IGF-I. Mediates insulin-stimulated protein synthesis by phosphorylating TSC2 at 'Ser-939' and 'Thr-1462', thereby activating mTORC1 signaling and leading to both phosphorylation of 4E-BP1 and in activation of RPS6KB1. Promotes glycogen synthesis by mediating the insulin-induced activation of glycogen synthase. The activated form can suppress FoxO gene transcription and promote cell cycle progression. Essential for the SPATA13-mediated regulation of cell migration and adhesion assembly and disassembly.
  • Tissue specificity

    Expressed in all human cell types so far analyzed. The Tyr-176 phosphorylated form shows a significant increase in expression in breast cancers during the progressive stages i.e. normal to hyperplasia (ADH), ductal carcinoma in situ (DCIS), invasive ductal carcinoma (IDC) and lymph node metastatic (LNMM) stages.
  • Involvement in disease

    Defects in AKT1 are a cause of susceptibility to breast cancer (BC) [MIM:114480]. A common malignancy originating from breast epithelial tissue. Breast neoplasms can be distinguished by their histologic pattern. Invasive ductal carcinoma is by far the most common type. Breast cancer is etiologically and genetically heterogeneous. Important genetic factors have been indicated by familial occurrence and bilateral involvement. Mutations at more than one locus can be involved in different families or even in the same case.
    Defects in AKT1 are associated with colorectal cancer (CRC) [MIM:114500].
    Defects in AKT1 are associated with susceptibility to ovarian cancer [MIM:604370]; also called susceptibility to familial breast-ovarian cancer type 1 (BROVCA1).
  • 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. The PH domain mediates interaction with TNK2 and Tyr-176 is also essential for this interaction.
    The AGC-kinase C-terminal mediates interaction with THEM4.
  • Post-translational
    modifications

    Phosphorylation on Thr-308, Ser-473 and Tyr-474 is required for full activity. Activated TNK2 phosphorylates it on Tyr-176 resulting in its binding to the anionic plasma membrane phospholipid PA. This phosphorylated form localizes to the cell membrane, where it is targeted by PDPK1 and PDPK2 for further phosphorylations on Thr-308 and Ser-473 leading to its activation. Ser-473 phosphorylation by mTORC2 favors Thr-308 phosphorylation by PDPK1. Ser-473 phosphorylation is enhanced by interaction with AGAP2 isoform 2 (PIKE-A). Ser-473 phosphorylation is enhanced in focal cortical dysplasias with Taylor-type balloon cells.
    Ubiquitinated; undergoes both 'Lys-48'- and 'Lys-63'-linked polyubiquitination. TRAF6-induced 'Lys-63'-linked AKT1 ubiquitination is critical for phosphorylation and activation. When ubiquitinated, it translocates to the plasma membrane, where it becomes phosphorylated. 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. Nucleus. Cell membrane. Nucleus after activation by integrin-linked protein kinase 1 (ILK1). Nuclear translocation is enhanced by interaction with TCL1A. Phosphorylation on Tyr-176 by TNK2 results in its localization to the cell membrane where it is targeted for further phosphorylations on Thr-308 and Ser-473 leading to its activation and the activated form translocates to the nucleus.
  • Target information above from: UniProt accession P31749 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 207 Human
    • Entrez Gene: 11651 Mouse
    • Entrez Gene: 24185 Rat
    • Omim: 164730 Human
    • SwissProt: P31749 Human
    • SwissProt: P31750 Mouse
    • SwissProt: P47196 Rat
    • Unigene: 525622 Human
    • Unigene: 6645 Mouse
    • Unigene: 11422 Rat
    see all
  • Alternative names

    • AKT 1 antibody
    • AKT antibody
    • AKT1 antibody
    • AKT1_HUMAN antibody
    • C AKT antibody
    • cAKT antibody
    • MGC99656 antibody
    • PKB alpha antibody
    • PKB antibody
    • PKB-ALPHA antibody
    • PRKBA antibody
    • Protein Kinase B Alpha antibody
    • Protein kinase B antibody
    • Proto-oncogene c-Akt antibody
    • RAC Alpha antibody
    • RAC antibody
    • Rac protein kinase alpha antibody
    • RAC Serine/Threonine Protein Kinase antibody
    • RAC-alpha serine/threonine-protein kinase antibody
    • RAC-PK-alpha antibody
    • v akt murine thymoma viral oncogene homolog 1 antibody
    • vAKT Murine Thymoma Viral Oncogene Homolog 1 antibody
    see all

Images

  • Western blot - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)
    Western blot - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)
    All lanes : Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283) at 1/25000 dilution

    Lane 1 : NIH/3T3 (Mouse embryonic fibroblast) treated with 100ng/ml PDGF for 1 hour whole cell lysates
    Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates

    Lysates/proteins at 15 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: 3 minutes


    Blocking buffer and concentration: 5% NFDM/TBST

  • Western blot - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)
    Western blot - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)
    All lanes : Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283) 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)

    Predicted band size: 56 kDa


    Exposure time: 50 seconds


    The 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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)

    ab81283, at 1/100 dilution, staining AKT1 in untreated (left panel) and Phosphatase-treated (right panel) human cervical carcinoma by Immunohistochemistry using formalin-fixed, paraffin-embedded tissue

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Western blot - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)
    Western blot - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)
    All lanes : Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283) at 0.259 µg/ml

    Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) grown in serum free media overnight, whole cell lysate
    Lane 2 : HeLa grown in serum free media overnight, then treated with 150nM Insulin for 5min, whole cell lysate
    Lane 3 : HeLa grown in serum free media overnight, then treated with 150nM Insulin for 5min, whole cell lysate. Then the membrane was incubated with alkaline phosphatase

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 56 kDa



    Blocking and diluting buffer: 5% NFDM/TBST. 

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)Image from Henken FE et al., Mol Cancer. 2011 Jun 10;10:71. Fig 7.; doi:10.1186/1476-4598-10-71; 10 June 2011, Molecular Cancer 2011, 10:71

    Immunohistochemical analysis of Human HPV16 immortalized keratinocytes transfected with non-targeting siRNA, staining AKT1 (phospho S473) (green) with ab81283.
    Antigen retrieval was performed by heat mediation in citrate buffer (pH 6). Samples were blocked with 10% goat serum before incubating with primary antibody (1/100). Fluoroscein-conjugated tyramide was used to detect staining.

  • Western blot - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)
    Western blot - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)

    MCF7 cells were incubated at 37°C for 2 hours with vehicle control (0 μM) and different concentrations of CCCP (ab 141229). Increased expression of AKT1 (phospho S473) (ab81283) in MCF7 cells correlates with an increase in CCCP concentration, as described in literature.

    Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 10μg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 5% BSA before being incubated with ab81283 at 2 μg/ml and ab8227 at 1 μg/ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (ab97051) at 1/10000 and visualised using ECL development solution.

  • Dot Blot - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)
    Dot Blot - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)

    Dot blot analysis of AKT1 (phospho S473) phospho peptide (Lane 1) and AKT1 non-phospho peptide (Lane 2) labelling AKT1 (phospho S473) phospho peptide with ab81283 at a dilution of 1:1000 dilution (0.259μg/ml). A Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) was used as the secondary antibody at a dilution of 1:20,000 dilution.

    Blocking buffer: 5% NFDM/TBST. Dilution buffer: 5% NFDM /TBST.

  • Western blot - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)
    Western blot - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)
    All lanes : Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283) at 0.259 µg/ml

    Lane 1 : LNCaP (human prostate carcinoma epithelial cell) whole cell lysate
    Lane 2 : LNCaP treated with 100nM Cacyculin A for 30min whole cell lysate
    Lane 3 : LNCaP treated with 100nM Cacyculin A for 30min whole cell lysate. Then the membrane was incubated with alkaline phosphatase

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 56 kDa



    Blocking and diluting buffer: 5% NFDM/TBST.

  • Western blot - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)
    Western blot - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)
    All lanes : Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283) at 1/1000 dilution

    All lanes : HEK293 cell lysate

    Predicted band size: 56 kDa



    Insulin treatment: cells were starved overnight and then treated for 20 min (Insulin) at 100 ng/ml.

    Phosphatase treatment: membrane strips were incubated with 200 ul of phosphatase (150 U/ml) at 37 degrees for 1 hour.

  • Western blot - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)
    Western blot - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)
    All lanes : Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283) at 1/1000 dilution

    All lanes : NIH/3T3 cell lysate

    Predicted band size: 56 kDa



    PDGF treatment: cells were starved overnight and then treated for 1 h with PDGF at 100 ng/ml.

    Phosphatase treatment: membrane strips were incubated with 200 ul of phosphatase (150 U/ml) at 37 degrees for 1 hour.

  • Western blot - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)
    Western blot - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)

    Primary : All Lanes : Anti AKT1 (phospho S473) antibody (ab81283) at 1:5000 dilution. Lane 1 = AKT1 (His tag) full length recombinant protein ab62279 - 50ng. Lane 2 = NIH3T3 serum starved overnight, 15ug. Lane 3 = NIH3T3 serum starved overnight and treated with PDGF-AB 50ng/mL for 1 hour, 15ug. Secondary : Lanes 1-3 : Goat polyclonal to Rabbit IgG H&L Pre-Adsorbed (HRP) at 1:5000 developed using the ECL technique. Performed under reducing conditions (50mM DTT, Sample heated at 60ºC). Predicted band size : 56kDa. Observed band size : 56kDa. Blocking step: 5% Milk in 50mM Tris+0.05% Tween for 1 hour at RT. Primary antibody buffer: 5% BSA in 50mM Tris+0.05% Tween overnight. Secondary antibody buffer: 5% Milk in 50mM Tris+0.05% Tween for 2 hours at RT. Exposure time : 5 minutes

  • In-Cell ELISA - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)
    In-Cell ELISA - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)
    NIH3T3 cells were starved overnight and treated with PDGF 50ng/mL or vehicle control for 1 hour prior to fixation with 4% paraformaldehyde. Levels of total Akt were measured using antibody ab81283 on an infrared in cell ELISA assay platform.
  • Western blot - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)
    Western blot - Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)

    PC12 cells were incubated at 37°C for 24 hours with vehicle control (0 nM) and 1 μM of Galanin (1-15) (porcine, rat) (ab 141152). Decreased expression of AKT1 (phospho S473) (ab81283) in PC12 cells correlates with an increase in Galanin (1-15) (porcine, rat) concentration, as described in literature.

     Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 30μg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 3% milk before being incubated withab81283at 1 μg/ml andab8227at 1 μg/ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (ab97051) at 1/10000 and visualised using ECL development solution.

  • Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)
    Anti-AKT1 (phospho S473) antibody [EP2109Y] (ab81283)

Protocols

  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (481)

Publishing research using ab81283? Please let us know so that we can cite the reference in this datasheet.

ab81283 has been referenced in 481 publications.

  • Fu J  et al. MicroRNA-451a prevents cutaneous squamous cell carcinoma progression via the 3-phosphoinositide-dependent protein kinase-1-mediated PI3K/AKT signaling pathway. Exp Ther Med 21:116 (2021). PubMed: 33335579
  • Li Z  et al. Kcnh2 mediates FAK/AKT-FOXO3A pathway to attenuate sepsis-induced cardiac dysfunction. Cell Prolif 54:e12962 (2021). PubMed: 33263944
  • Liu X  et al. Huaier shows anti-cancer activities by inhibition of cell growth, migration and energy metabolism in lung cancer through PI3K/AKT/HIF-1a pathway. J Cell Mol Med 25:2228-2237 (2021). PubMed: 33377619
  • Tao J  et al. Sfrp1 protects against acute myocardial ischemia (AMI) injury in aged mice by inhibiting the Wnt/ß-catenin signaling pathway. J Cardiothorac Surg 16:12 (2021). PubMed: 33468190
  • Chen D  et al. Suppressive effect of platycodin D on bladder cancer through microRNA-129-5p-mediated PABPC1/PI3K/AKT axis inactivation. Braz J Med Biol Res 54:e10222 (2021). PubMed: 33470388
View all Publications for this product

Customer reviews and Q&As

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1-10 of 11 Abreviews or Q&A

IHC - Wholemount abreview for Anti-AKT1 (phospho S473) antibody [EP2109Y]

Good
Abreviews
Abreviews
abreview image
Application
IHC - Wholemount
Sample
Mouse Tissue (cornea)
Specification
cornea
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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 May 24 2022

Western blot abreview for Anti-AKT1 (phospho S473) antibody [EP2109Y]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (adipocytes 3T3-L1)
Gel Running Conditions
Reduced Denaturing (Nupage Bis-Tris Glycine gradient precast gel)
Loading amount
30 µg
Treatment
High glucose and inflammatory conditions
Specification
adipocytes 3T3-L1
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.

PROF. Michele D'angelo

Verified customer

Submitted Sep 29 2021

Immunocytochemistry/ Immunofluorescence abreview for Anti-AKT1 (phospho S473) antibody [EP2109Y]

Excellent
Abreviews
Abreviews
This product is known to not work in this application or species.
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (GBM cell line)
Permeabilization
Yes - cold methanol
Specification
GBM cell line
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5 · Temperature: 23°C
Fixative
Formaldehyde
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.

Miss. nomi zalcman

Verified customer

Submitted Sep 17 2020

Immunocytochemistry/ Immunofluorescence abreview for Anti-AKT1 (phospho S473) antibody [EP2109Y]

Good
Abreviews
Abreviews
This product is known to not work in this application or species.
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Bos taurus Cell (Ovarian follicle, granulosa cells)
Permeabilization
Yes - 0.1% PBST
Specification
Ovarian follicle, granulosa cells
Blocking step
(agent) for 1 hour(s) and 0 minute(s) · Concentration: 20% · Temperature: 23°C
Fixative
Paraformaldehyde
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 18 2020

Western blot abreview for Anti-AKT1 (phospho S473) antibody [EP2109Y]

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK 293 cells)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
20 µg
Specification
HEK 293 cells
Blocking step
BSA 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 Sep 23 2016

Western blot abreview for Anti-AKT1 (phospho S473) antibody [EP2109Y]

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Epithelial OVCA cell line)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
100 µg
Treatment
0- 200µM Perifosine 24 hr
Specification
Epithelial OVCA cell line
Blocking step
Milk as blocking agent for 12 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 Jul 13 2016

Immunocytochemistry/ Immunofluorescence abreview for Anti-AKT1 (phospho S473) antibody [EP2109Y]

Excellent
Abreviews
Abreviews
This product is known to not work in this application or species.
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Peritonei tumor)
Permeabilization
Yes - 0.3%Triton X 100
Specification
Peritonei tumor
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Paraformaldehyde
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 Jan 19 2016

Western blot abreview for Anti-AKT1 (phospho S473) antibody [EP2109Y]

Average
Abreviews
Abreviews
Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing
Sample
Human Cell lysate - whole cell (macrophage)
Specification
macrophage
Treatment
0.1 ug/ml LPS 15 min
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°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 Dec 12 2014

Immunohistochemistry (Frozen sections) abreview for Anti-AKT1 (phospho S473) antibody [EP2109Y]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Sample
Human Cell (Peritoneal tumor)
Specification
Peritoneal tumor
Permeabilization
No
Fixative
Paraformaldehyde
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 Oct 14 2014

Western blot abreview for Anti-AKT1 (phospho S473) [EP2109Y] antibody

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (4-12%)
Sample
Human Cell lysate - whole cell (A549)
Specification
A549
Treatment
Serum starvation followed by serum addition together with 100ng/ml of EGF for 0 to 3hrs
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°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 Mar 21 2014

1-10 of 11 Abreviews or Q&A

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