Recombinant
RabMAb

Recombinant Anti-AKT1 + AKT2 + AKT3 (phospho S472 + S473 + S474) antibody [EPR18853] - BSA and Azide free (ab222489)

Overview

  • Product name

    Anti-AKT1 + AKT2 + AKT3 (phospho S472 + S473 + S474) antibody [EPR18853] - BSA and Azide free
    See all AKT1 + AKT2 + AKT3 primary antibodies
  • Description

    Rabbit monoclonal [EPR18853] to AKT1 + AKT2 + AKT3 (phospho S472 + S473 + S474) - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IF, IP, Dot blotmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human AKT1 + AKT2 + AKT3 aa 450 to the C-terminus (phospho S472). The exact sequence is proprietary.
    Database link: Q9Y243

  • Positive control

    • WB: MCF7 whole cell lysate treated with 100ng/ml IGF-1 for 15 minutes; PC-12 and NIH/3T3 whole cell lysates treated with 100ng/ml PDGF for 60 minutes. ICC/IF: NIH/3T3 cells treated with PDGF (100 ng/ml) for 1 hour. IP: NIH/3T3 treated with 50ng/ml PDGF for 40min whole cell lysate.
  • General notes

    Ab222489 is the carrier-free version of ab192623. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab222489 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

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

    This product is a recombinant rabbit monoclonal antibody.

Applications

Our Abpromise guarantee covers the use of ab222489 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 Use at an assay dependent concentration. Detects a band of approximately 56 kDa (predicted molecular weight: 56 kDa).
ICC/IF Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
Dot blot Use at an assay dependent concentration.

Target

  • Function

    IGF-1 leads to the activation of AKT3, which may play a role in regulating cell survival. Capable of phosphorylating several known proteins. Truncated isoform 2/PKB gamma 1 without the second serine phosphorylation site could still be stimulated but to a lesser extent.
  • Tissue specificity

    In adult tissues, it is highly expressed in brain, lung and kidney, but weakly in heart, testis and liver. In fetal tissues, it is highly expressed in heart, liver and brain and not at all in kidney.
  • 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.
  • Post-translational
    modifications

    Phosphorylation on Thr-305 and Ser-472 is required for full activity (By similarity). Phosphorylated upon DNA damage, probably by ATM or ATR.
    Ubiquitinated. 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. Membrane. Membrane-associated after cell stimulation leading to its translocation.
  • Information by UniProt
  • Database links

  • Alternative names

    • AKT antibody
    • AKT1 antibody
    • AKT1 kinase antibody
    • AKT1m antibody
    • AKT2 antibody
    • AKT2 kinase antibody
    • Akt3 antibody
    • AKT3_HUMAN antibody
    • CAKT antibody
    • CWS6 antibody
    • DKFZp434N0250 antibody
    • HIHGHH antibody
    • kinase Akt1 antibody
    • MGC99656 antibody
    • MPPH antibody
    • Murine thymoma viral (v-akt) homolog 2 antibody
    • pan-akt antibody
    • PKB ALPHA antibody
    • PKB antibody
    • PKB beta antibody
    • PKB gamma antibody
    • PKB-GAMMA antibody
    • PKB/Akt antibody
    • PKBALPHA antibody
    • PKBB antibody
    • PKBBETA antibody
    • PKBG antibody
    • PKBGAMMA antibody
    • PRKBA antibody
    • PRKBB antibody
    • PRKBG antibody
    • Protein kinase Akt 2 antibody
    • Protein kinase Akt-2 antibody
    • Protein kinase Akt-3 antibody
    • Protein kinase B alpha antibody
    • Protein kinase B antibody
    • Protein kinase B beta antibody
    • Protein kinase B gamma antibody
    • Proto oncogene c Akt antibody
    • Proto-oncogene c-Akt antibody
    • RAC ALPHA antibody
    • RAC alpha serine/threonine protein kinase antibody
    • RAC antibody
    • RAC BETA antibody
    • RAC beta serine/threonine protein kinase antibody
    • RAC PK alpha antibody
    • RAC PK beta antibody
    • rac protein kinase alpha antibody
    • rac protein kinase beta antibody
    • RAC-ALPHA antibody
    • RAC-alpha serine/threonine-protein kinase antibody
    • RAC-beta serine/threonine-protein kinase antibody
    • RAC-gamma antibody
    • RAC-gamma serine/threonine-protein kinase antibody
    • RAC-PK-alpha antibody
    • RAC-PK-beta antibody
    • RAC-PK-gamma antibody
    • RACALPHA antibody
    • RACalpha serine/threonine kinase antibody
    • RACBETA antibody
    • RACgamma antibody
    • RACgamma serine/threonine protein kinase antibody
    • RACPKgamma antibody
    • serine threonine protein kinase antibody
    • STK 2 antibody
    • STK-2 antibody
    • STK2 antibody
    • thymoma viral proto oncogene 1 antibody
    • thymoma viral proto oncogene antibody
    • V akt murine thymoma viral oncogene homolog 1 antibody
    • V akt murine thymoma viral oncogene homolog 2 antibody
    • V akt murine thymoma viral oncogene homolog 3 (protein kinase B, gamma) antibody
    • V akt murine thymoma viral oncogene homolog 3 antibody
    • V-AKT murine thymoma viral oncogene homolog 1 antibody
    • V-AKT murine thymoma viral oncogene homolog 2 antibody
    • V-AKT murine thymoma viral oncogene homolog 3 antibody
    • vakt murine thymoma viral oncogene homolog 1 antibody
    • vakt murine thymoma viral oncogene homolog 2 antibody
    • vakt murine thymoma viral oncogene homolog 3 antibody
    see all

Images

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells, untreated or treated with PDGF (100 ng/ml) for 1 hour, labeling AKT3 (phospho S472) + AKT2 (phospho S474) + AKT1 (phospho S473) with ab192623 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    The signal increased after treatment with PDGF (100 ng/ml) for 1 hour on NIH/3T3 cells.

    The nuclear counter stain is DAPI (blue).

    Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab192623).

  • AKT3 (phospho S472) was immunoprecipitated from 0.35 mg of NIH/3T3 (Mouse embryonic fibroblast cell line) treated with 50ng/ml PDGF for 40min whole cell lysate with ab192623 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab192623 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

    Lane 1: NIH/3T3 treated with 50ng/ml PDGF for 40min whole cell lysate, 10μg (Input).

    Lane 2: ab192623 IP in NIH/3T3 treated with 50ng/ml PDGF for 40min whole cell lysate.

    Lane 3: Rabbit IgG,monoclonal [EPR25A]-Isotype Control (ab172730) instead of ab192623 in NIH/3T3 treated with 50ng/ml PDGF for 40min whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 5 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab192623).

  • Dot blot analysis of AKT3 (phospho S472) labeled with ab192623 at 1/1000 dilution.

    Lane 1: AKT3 (phospho S472) phospho peptide;

    Lane 2: AKT3 non-phospho peptide;

    Lane 3: AKT1 (phospho S473) phospho peptide;

    Lane 4: AKT2 (phospho S474) phospho peptide.

    Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated (ab97051) at 1/100000 dilution was used as secondary antibody.

    Blocking and diluting buffer: 5% NFDM/TBST.

    Exposure time: 3 minutes.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab192623).

References

ab222489 has not yet been referenced specifically in any publications.

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