Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-PTEN antibody [EPR4408-76] (ab133532)

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Western blot - Anti-PTEN antibody [EPR4408-76] (ab133532)
  • Western blot - Anti-PTEN antibody [EPR4408-76] (ab133532)
  • Western blot - Anti-PTEN antibody [EPR4408-76] (ab133532)
  • Western blot - Anti-PTEN antibody [EPR4408-76] (ab133532)
  • Anti-PTEN antibody [EPR4408-76] (ab133532)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR4408-76] to PTEN
  • Suitable for: WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-PTEN antibody [EPR4408-76]
    See all PTEN primary antibodies

  • Description

    Rabbit monoclonal [EPR4408-76] to PTEN

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WBmore details
    Unsuitable for: Flow Cyt,ICC/IF,IHC-P or IP

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant full length protein corresponding to Human PTEN.

  • Positive control

    • WB: Hap1, C6, RAW 264.7, NIH 3T3, MCF7, 293T, A431, and HeLa cell lysates.

  • 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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab133532 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 (1)
1/10000 - 1/50000. Predicted molecular weight: 47 kDa.
Notes
WB
1/10000 - 1/50000. Predicted molecular weight: 47 kDa.
  • Application notes
    Is unsuitable for Flow Cyt,ICC/IF,IHC-P or IP.

    • Target

    • Function

      Tumor suppressor. Acts as a dual-specificity protein phosphatase, dephosphorylating tyrosine-, serine- and threonine-phosphorylated proteins. Also acts as a lipid phosphatase, removing the phosphate in the D3 position of the inositol ring from phosphatidylinositol 3,4,5-trisphosphate, phosphatidylinositol 3,4-diphosphate, phosphatidylinositol 3-phosphate and inositol 1,3,4,5-tetrakisphosphate with order of substrate preference in vitro PtdIns(3,4,5)P3 > PtdIns(3,4)P2 > PtdIns3P > Ins(1,3,4,5)P4. The lipid phosphatase activity is critical for its tumor suppressor function. Antagonizes the PI3K-AKT/PKB signaling pathway by dephosphorylating phosphoinositides and thereby modulating cell cycle progression and cell survival. The unphosphorylated form cooperates with AIP1 to suppress AKT1 activation. Dephosphorylates tyrosine-phosphorylated focal adhesion kinase and inhibits cell migration and integrin-mediated cell spreading and focal adhesion formation. 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. May be a negative regulator of insulin signaling and glucose metabolism in adipose tissue. The nuclear monoubiquitinated form possesses greater apoptotic potential, whereas the cytoplasmic nonubiquitinated form induces less tumor suppressive ability. In motile cells, suppresses the formation of lateral pseudopods and thereby promotes cell polarization and directed movement.
      Isoform alpha: Functional kinase, like isoform 1 it antagonizes the PI3K-AKT/PKB signaling pathway. Plays a role in mitochondrial energetic metabolism by promoting COX activity and ATP production, via collaboration with isoform 1 in increasing protein levels of PINK1.

    • Tissue specificity

      Expressed at a relatively high level in all adult tissues, including heart, brain, placenta, lung, liver, muscle, kidney and pancreas.

    • Involvement in disease

      Cowden syndrome 1
      Lhermitte-Duclos disease
      Bannayan-Riley-Ruvalcaba syndrome
      Squamous cell carcinoma of the head and neck
      Endometrial cancer
      PTEN mutations are found in a subset of patients with Proteus syndrome, a genetically heterogeneous condition. The molecular diagnosis of PTEN mutation positive cases classifies Proteus syndrome patients as part of the PTEN hamartoma syndrome spectrum. As such, patients surviving the early years of Proteus syndrome are likely at a greater risk of developing malignancies.
      Glioma 2
      VACTERL association with hydrocephalus
      Prostate cancer
      Macrocephaly/autism syndrome
      A microdeletion of chromosome 10q23 involving BMPR1A and PTEN is a cause of chromosome 10q23 deletion syndrome, which shows overlapping features of the following three disorders: Bannayan-Zonana syndrome, Cowden disease and juvenile polyposis syndrome.

    • Sequence similarities

      Contains 1 C2 tensin-type domain.
      Contains 1 phosphatase tensin-type domain.

    • Domain

      The C2 domain binds phospholipid membranes in vitro in a Ca(2+)-independent manner; this binding is important for its tumor suppressor function.

    • Post-translational
      modifications

      Constitutively phosphorylated by CK2 under normal conditions. Phosphorylated in vitro by MAST1, MAST2, MAST3 and STK11. Phosphorylation results in an inhibited activity towards PIP3. Phosphorylation can both inhibit or promote PDZ-binding. Phosphorylation at Tyr-336 by FRK/PTK5 protects this protein from ubiquitin-mediated degradation probably by inhibiting its binding to NEDD4. Phosphorylation by ROCK1 is essential for its stability and activity. Phosphorylation by PLK3 promotes its stability and prevents its degradation by the proteasome.
      Monoubiquitinated; monoubiquitination is increased in presence of retinoic acid. Deubiquitinated by USP7; leading to its nuclear exclusion. Monoubiquitination of one of either Lys-13 and Lys-289 amino acid is sufficient to modulate PTEN compartmentalization. Ubiquitinated by XIAP/BIRC4.

    • Cellular localization

      Secreted. May be secreted via a classical signal peptide and reenter into cells with the help of a poly-Arg motif and Cytoplasm. Nucleus. Nucleus, PML body. Monoubiquitinated form is nuclear. Nonubiquitinated form is cytoplasmic. Colocalized with PML and USP7 in PML nuclear bodies. XIAP/BIRC4 promotes its nuclear localization.
    • Information by UniProt

    • Database links

      see all

    • Alternative names

      • 10q23del antibody
      • BZS antibody
      • DEC antibody
      • GLM2 antibody
      • MGC11227 antibody
      • MHAM antibody
      • MMAC1 antibody
      • MMAC1 phosphatase and tensin homolog deleted on chromosome 10 antibody
      • Mutated in multiple advanced cancers 1 antibody
      • Phosphatase and tensin homolog antibody
      • Phosphatase and tensin like protein antibody
      • Phosphatidylinositol 3,4,5-trisphosphate 3-phosphatase and dual-specificity protein phosphatase PTEN antibody
      • Pten antibody
      • PTEN_HUMAN antibody
      • PTEN1 antibody
      • TEP1 antibody
      see all

    Images

    • Western blot - Anti-PTEN antibody [EPR4408-76] (ab133532)
      Western blot - Anti-PTEN antibody [EPR4408-76] (ab133532)
      All lanes : Anti-PTEN antibody [EPR4408-76] (ab133532) at 1/10000 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : PTEN knockout HeLa cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 47 kDa
      Observed band size: 47 kDa



      Lanes 1- 2: Merged signal (red and green). Green - ab133532 observed at 47 kDa. Red - Anti-Vinculin antibody [VIN-54] observed at 124 kDa.

       ab133532 was shown to react with PTEN in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255419 (knockout cell lysate ab263829) was used. Wild-type HeLa and PTEN knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab133532 and Anti-Vinculin antibody [VIN-54] overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-PTEN antibody [EPR4408-76] (ab133532)
      Western blot - Anti-PTEN antibody [EPR4408-76] (ab133532)
      All lanes : Anti-PTEN antibody [EPR4408-76] (ab133532) at 1/10000 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : PTEN CRISPR/Cas9 edited HeLa cell lysate
      Lane 3 : HAP1 cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 47 kDa
      Observed band size: 47 kDa



      Lanes 1 - 3: Merged signal (red and green). Green - ab133532 observed at 47 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

      ab133532 was shown to react with PTEN in wild-type HeLa cells in western blot. The band observed in PTEN CRISPR/Cas9 edited cell line ab255419 (PTEN CRISPR/Cas9 edited cell lysate ab263829) below 47 kDa may represent truncated forms and cleaved fragments. This has not been investigated further. HeLa wild-type and PTEN CRISPR/Cas9 edited cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab133532 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

    • Western blot - Anti-PTEN antibody [EPR4408-76] (ab133532)
      Western blot - Anti-PTEN antibody [EPR4408-76] (ab133532)
      Lanes 1-2 : Anti-PTEN antibody [EPR4408-76] (ab133532) at 1/10000 dilution
      Lanes 3-4 : Anti-GAPDH antibody [6C5] - Loading Control (ab8245) at 1/2000 dilution


      Lanes 1 & 3 & 5 : PTEN knockout HAP1 cell lysate
      Lanes 2 & 4 & 6 : Wild-type HAP1 cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 47 kDa



      Lanes 1 and 2: Green signal from target - ab133532 observed at 47 kDa
      Lanes 3 and 4: Red signal from loading control - ab8245 observed at 37 kDa
      Lanes 5 and 6: Merged (red and green) signal


      ab133532 was shown to specifically react with PTEN in wild-type HAP1 cells. No band was observed when PTEN knockout samples were used. Wild-type and PTEN knockout samples were subjected to SDS-PAGE, ab133532 and ab8245 (loading control to GAPDH) were diluted to 1/10,000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1hr at room temperature before imaging.

    • Western blot - Anti-PTEN antibody [EPR4408-76] (ab133532)
      Western blot - Anti-PTEN antibody [EPR4408-76] (ab133532)
      All lanes : Anti-PTEN antibody [EPR4408-76] (ab133532) at 1/10000 dilution

      Lane 1 : C6 cell lysate
      Lane 2 : RAW 264.7 cell lysate
      Lane 3 : NIH 3T3 cell lysate
      Lane 4 : MCF7 cell lysate
      Lane 5 : 293T cell lysate
      Lane 6 : A431 cell lysate
      Lane 7 : HeLa cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size: 47 kDa
      Observed band size: 54 kDa why is the actual band size different from the predicted?

    • Anti-PTEN antibody [EPR4408-76] (ab133532)
      Anti-PTEN antibody [EPR4408-76] (ab133532)

    References (2)

    Publishing research using ab133532? Please let us know so that we can cite the reference in this datasheet.

    ab133532 has been referenced in 2 publications.

    • Sikorski K  et al. A high-throughput pipeline for validation of antibodies. Nat Methods 15:909-912 (2018). PubMed: 30377371
    • Zhong C  et al. miR-19b controls cardiac fibroblast proliferation and migration. J Cell Mol Med 20:1191-7 (2016). WB . PubMed: 27061862

    Customer reviews and Q&As

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    Western blot abreview for Anti-PTEN antibody [EPR4408-76]

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HEK293)
    Gel Running Conditions
    Reduced Denaturing (12%)
    Loading amount
    20 µg
    Specification
    HEK293
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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    Submitted Mar 04 2021

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