Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-PTEN antibody [EPR22636-122] (ab267787)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR22636-122] to PTEN
  • Suitable for: WB, IHC-P, Flow Cyt, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-PTEN antibody [EPR22636-122]
    See all PTEN primary antibodies

  • Description

    Rabbit monoclonal [EPR22636-122] to PTEN

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt, IPmore details
    Unsuitable for: ICC/IF

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human PTEN aa 350 to the C-terminus. The exact sequence is proprietary.
    Database link: P60484

  • Positive control

    • WB: Wild-type HAP1, MCF7, MDA-MB-468, HeLa, C6, RAW264.7, PC-12 and NIH/3T3 whole cell lysates; Mouse kidney, Mouse spleen and Rat lung lysates. IHC-P: Human endometrial cancer, Human ovarian cancer, Human pancreas, Mouse pancreas and Rat pancreas tissues. Flow Cyt: HeLa and NIH/3T3 cells. IP: MCF7 and HeLa cells.

  • 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

Our Abpromise guarantee covers the use of ab267787 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 1/1000. Detects a band of approximately 54 kDa (predicted molecular weight: 47 kDa).
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt 1/500.
IP 1/30.
  • Application notes
    Is unsuitable for ICC/IF.

    • 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 [EPR22636-122] (ab267787)
      Western blot - Anti-PTEN antibody [EPR22636-122] (ab267787)
      All lanes : Anti-PTEN antibody [EPR22636-122] (ab267787) at 1/1000 dilution

      Lane 1 : C6 (rat glial tumor glial cell), whole cell lysate
      Lane 2 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate
      Lane 3 : PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate
      Lane 4 : NIH/3T3 (mouse embryonic fibroblast), whole cell lysate
      Lane 5 : Mouse kidney tissue lysate
      Lane 6 : Mouse spleen tissue lysate
      Lane 7 : Rat lung tissue lysate

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



      Blocking and dilution buffer: 5% NFDM/TBST.

      Exposure times:

      Lanes 1-6: 3 seconds;
      Lane 7: 7 seconds.

      The molecular weight observed is consistent with what has been described in the literature (PMID:10514407).

    • Western blot - Anti-PTEN antibody [EPR22636-122] (ab267787)
      Western blot - Anti-PTEN antibody [EPR22636-122] (ab267787)
      All lanes : Anti-PTEN antibody [EPR22636-122] (ab267787) at 1/1000 dilution

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : PTEN knockout HAP1 whole cell lysate
      Lane 3 : MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate
      Lane 4 : MDA-MB-468 (human breast adenocarcinoma epithelial cell), whole cell lysate
      Lane 5 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate

      Lysates/proteins at 20 µg per lane.

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

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



      Blocking and dilution buffer: 5% NFDM/TBST.

      Exposure times:

      Lanes 1-4: 3 seconds;
      Lane 5: 114 seconds.

      ab267787 was shown to specifically react with PTEN in wild-type HAP1 cells as signal was lost in PTEN knockout cells. Wild-type and PTEN knockout samples were subjected to SDS-PAGE. Ab267787 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/20,000 dilution for 1 hour at room temperature before imaging.

      The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID:10514407).

      Negative control: MDA-MB-468 (PMID:21358673,15674339).

       

       

    • Immunoprecipitation - Anti-PTEN antibody [EPR22636-122] (ab267787)
      Immunoprecipitation - Anti-PTEN antibody [EPR22636-122] (ab267787)

      PTEN was immunoprecipitated from 0.35 mg HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate with ab267787 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab267787 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

      Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10ug

      Lane 2: ab267787 IP in HeLa whole cell lysate

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab267787 in HeLa whole cell lysate

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

      Exposure time: 15 seconds

       

    • Immunoprecipitation - Anti-PTEN antibody [EPR22636-122] (ab267787)
      Immunoprecipitation - Anti-PTEN antibody [EPR22636-122] (ab267787)

      PTEN was immunoprecipitated from 0.35 mg MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate with ab267787 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab267787 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

      Lane 1: MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate 10ug

      Lane 2: ab267787 IP in MCF7 whole cell lysate

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab267787 in MCF7 whole cell lysate

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

      Exposure time: 15 seconds

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [EPR22636-122] (ab267787)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [EPR22636-122] (ab267787)

      Immunohistochemical analysis of paraffin-embedded Rat pancreas tissue labeling PTEN with ab267787 at 1/2000 dilution (2.18 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat pancreas (PMID:11021813). The section was incubated with ab267787 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [EPR22636-122] (ab267787)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [EPR22636-122] (ab267787)

      Immunohistochemical analysis of paraffin-embedded Mouse pancreas tissue labeling PTEN with ab267787 at 1/2000 dilution (2.18 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse pancreas (PMID:11021813). The section was incubated with ab267787 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins. 

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [EPR22636-122] (ab267787)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [EPR22636-122] (ab267787)

      Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling PTEN with ab267787 at 1/2000 dilution (2.18 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human pancreas (PMID:11021813). The section was incubated with ab267787 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [EPR22636-122] (ab267787)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [EPR22636-122] (ab267787)

      Immunohistochemical analysis of paraffin-embedded Human ovarian cancer tissue labeling PTEN with ab267787 at 1/2000 dilution (2.18 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on stroma of human ovarian cancer (PMID:25608477). The section was incubated with ab267787 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [EPR22636-122] (ab267787)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTEN antibody [EPR22636-122] (ab267787)

      Immunohistochemical analysis of paraffin-embedded Human endometrial cancer tissue labeling PTEN with ab267787 at 1/2000 dilution (2.18 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on stroma of human endometrial cancer (PMID:2230170). The section was incubated with ab267787 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    • Flow Cytometry - Anti-PTEN antibody [EPR22636-122] (ab267787)
      Flow Cytometry - Anti-PTEN antibody [EPR22636-122] (ab267787)

      Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast) cells labelling PTEN with ab267787 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

    • Flow Cytometry - Anti-PTEN antibody [EPR22636-122] (ab267787)
      Flow Cytometry - Anti-PTEN antibody [EPR22636-122] (ab267787)

      Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling PTEN with ab267787 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

    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

    References

    Publishing research using ab267787? Please let us know so that we can cite the reference in this datasheet.

    ab267787 has not yet been referenced specifically in any publications.

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