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    akt3--akt2--akt1-antibody-y89-ab32505.pdf

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

Recombinant Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)

  • Datasheet
  • SDS
Reviews (6)Q&A (1)References (126)

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Western blot - Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)
  • Immunocytochemistry/ Immunofluorescence - Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)
  • Immunoprecipitation - Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)
  • Western blot - Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)
  • Western blot - Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)
  • Flow Cytometry (Intracellular) - Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)
  • Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [Y89] to AKT3 + AKT2 + AKT1
  • Suitable for: Flow Cyt (Intra), ICC/IF, WB, IHC-P, IP
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free

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Overview

  • Product name

    Anti-AKT3 + AKT2 + AKT1 antibody [Y89]
  • Description

    Rabbit monoclonal [Y89] to AKT3 + AKT2 + AKT1
  • Host species

    Rabbit
  • Specificity

    This product reacts with AKT1, AKT2 and AKT3.
  • Tested applications

    Suitable for: Flow Cyt (Intra), ICC/IF, WB, IHC-P, IPmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat, Cow
  • Immunogen

    Synthetic peptide within Human AKT1 aa 450 to the C-terminus (C terminal). The exact sequence is proprietary.
    Database link: P31749

  • Positive control

    • MCF7 cell lysate and prostate carcinoma tissue. IP: MCF7 cell lysate
  • General notes

    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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    Y89
  • 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
    • Cardiovascular
    • Atherosclerosis
    • Diabetes associated
    • Cardiovascular
    • Heart
    • Cardiac metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Apoptosis
    • Metabolism
    • Types of disease
    • Diabetes
    • Metabolism
    • Types of disease
    • Heart disease
    • Cancer
    • Cell Death
    • Apoptosis
    • Metabolism

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab204252)
    • Alexa Fluor® 647 Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab204285)
    • Anti-AKT3 + AKT2 + AKT1 antibody [Y89] - BSA and Azide free (ab219588)
  • 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)
    • Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Related Products

    • Alsterpaullone, cyclin-dependent kinase (CDK) and GSK-3beta inhibitor (ab141070)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab32505 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
Flow Cyt (Intra)
1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF
1/100 - 1/250.
WB (3)
1/2000 - 1/10000. Detects a band of approximately 59 kDa (predicted molecular weight: 56 kDa).
IHC-P (1)
1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IP
1/50.
Notes
Flow Cyt (Intra)
1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF
1/100 - 1/250.
WB
1/2000 - 1/10000. Detects a band of approximately 59 kDa (predicted molecular weight: 56 kDa).
IHC-P
1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IP
1/50.

Target

  • Cellular localization

    AKT3: Cytoplasm. Membrane. Membrane-associated after cell stimulation leading to its translocation. AKT1: 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.
  • Database links

    • Entrez Gene: 100137872 Cow
    • Entrez Gene: 280991 Cow
    • Entrez Gene: 534923 Cow
    • Entrez Gene: 10000 Human
    • Entrez Gene: 207 Human
    • Entrez Gene: 208 Human
    • Entrez Gene: 11651 Mouse
    • Entrez Gene: 11652 Mouse
    • Entrez Gene: 23797 Mouse
    • Entrez Gene: 24185 Rat
    • Entrez Gene: 25233 Rat
    • Entrez Gene: 29414 Rat
    • Omim: 164730 Human
    • Omim: 164731 Human
    • Omim: 611223 Human
    • SwissProt: Q01314 Cow
    • SwissProt: P31749 Human
    • SwissProt: P31751 Human
    • SwissProt: Q9Y243 Human
    • SwissProt: P31750 Mouse
    • SwissProt: Q60823 Mouse
    • SwissProt: Q9WUA6 Mouse
    • SwissProt: P47196 Rat
    • SwissProt: P47197 Rat
    • SwissProt: Q63484 Rat
    • Unigene: 498292 Human
    • Unigene: 525622 Human
    • Unigene: 631535 Human
    • Unigene: 177194 Mouse
    • Unigene: 235194 Mouse
    • Unigene: 6645 Mouse
    • Unigene: 10506 Rat
    • Unigene: 11422 Rat
    • Unigene: 87066 Rat
    see all

Images

  • Western blot - Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)
    Western blot - Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)
    All lanes : Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505) at 1/10000 dilution

    Lane 1 : 293T cell lysate transfected with GFP tagged AKT1
    Lanes 2 & 4 : 293T cell lysate transfected with empty vector
    Lane 3 : 293T cell lysate transfected with GFP tagged AKT3

    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
    Observed band size: 82 kDa why is the actual band size different from the predicted?


    Exposure time: 8 seconds


    Blocking and diluting buffer and concentration: 5% NFDM/TBST

  • Immunocytochemistry/ Immunofluorescence - Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)
    Immunocytochemistry/ Immunofluorescence - Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)

    ab32505 staining in SK-N-SH cells treated with alsterpaullone (ab141070), by ICC/IF. Decrease of AKT1 + AKT2 + AKT3 expression correlates with increased concentration of alsterpaullone, as described in literature.
    The cells were incubated at 37°C for 6h in media containing different concentrations of ab141070 (alsterpaullone) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab32505 (1/200 dilution was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)

    Immunohistochemical analysis of paraffin-embedded prostate carcinoma using ab32505 at 1/100 dilution.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunoprecipitation - Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)
    Immunoprecipitation - Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)

    Purified ab32505 at 1/50 dilution (2µg) immunoprecipitating AKT3+AKT2+AKT1 in MCF7 whole cell lysate.
    Lane 1 (input): MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate 10µg
    Lane 2 (+): ab32505 + MCF7 whole cell lysate.
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32505 in MCF7 whole cell lysate.
    VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
    Blocking Buffer and concentration: 5% NFDM/TBST.
    Diluting buffer and concentration: 5% NFDM/TBST.
    Observed band size: 59 kDa

  • Western blot - Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)
    Western blot - Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)
    All lanes : Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505) at 1/2000 dilution

    Lane 1 : 293T cell lysate transfected with GFP tagged AKT2
    Lane 2 : 293T cell lysate transfected with empty vector

    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
    Observed band size: 82 kDa why is the actual band size different from the predicted?


    Exposure time: 5 seconds


    Blocking and diluting buffer and concentration: 5% NFDM/TBST

  • Western blot - Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)
    Western blot - Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)
    Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505) at 1/10000 dilution + MCF-7 cell lysate

    Predicted band size: 56 kDa
    Observed band size: 59 kDa why is the actual band size different from the predicted?

  • Flow Cytometry (Intracellular) - Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)
    Flow Cytometry (Intracellular) - Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)

    Overlay histogram showing HeLa cells stained with ab32505 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32505, 1/20 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a slightly decreased signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

     

  • Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)
    Anti-AKT3 + AKT2 + AKT1 antibody [Y89] (ab32505)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (126)

Publishing research using ab32505? Please let us know so that we can cite the reference in this datasheet.

ab32505 has been referenced in 126 publications.

  • Cui R  et al. AMOTL2 inhibits JUN Thr239 dephosphorylation by binding PPP2R2A to suppress the proliferation in non-small cell lung cancer cells. Biochim Biophys Acta Mol Cell Res 1868:118858 (2021). PubMed: 32950569
  • Meng X  et al. Bcl-2 Is Involved in Cardiac Hypertrophy through PI3K-Akt Pathway. Biomed Res Int 2021:6615502 (2021). PubMed: 33778070
  • 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
  • Cui H & Zhang Q Dexmedetomidine ameliorates lipopolysaccharide-induced acute lung injury by inhibiting the PI3K/Akt/FoxO1 signaling pathway. J Anesth 35:394-404 (2021). PubMed: 33821300
  • Gan D  et al. Repurposing of Pirfenidone (Anti-Pulmonary Fibrosis Drug) for Treatment of Rheumatoid Arthritis. Front Pharmacol 12:631891 (2021). PubMed: 33746759
View all Publications for this product

Customer reviews and Q&As

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1-3 of 3 Abreviews or Q&A

Western blot abreview for Anti-AKT1 antibody [Y89]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Loading amount
40 µg
Gel Running Conditions
Reduced Denaturing
Sample
Mouse Cell lysate - whole cell (Cardiomyocytes)
Specification
Cardiomyocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
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 11 2015

Western blot abreview for Anti-AKT1 [Y89] antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Loading amount
25 µg
Gel Running Conditions
Reduced Denaturing (4–15%)
Sample
Mouse Tissue lysate - whole (Lung)
Specification
Lung
Blocking step
BSA as blocking agent for 15 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 09 2014

Western blot abreview for Anti-AKT1 antibody [Y89]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Glioblastoma Initiating Cells (Primary cells))
Loading amount
30 µg
Specification
Glioblastoma Initiating Cells (Primary cells)
Gel Running Conditions
Reduced Denaturing (12.5)
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°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 Nov 16 2011

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