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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-PTEN antibody [EPR23729-4] (ab260011)

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Western blot - Anti-PTEN antibody [EPR23729-4] (ab260011)
  • Flow Cytometry (Intracellular) - Anti-PTEN antibody [EPR23729-4] (ab260011)
  • Immunoprecipitation - Anti-PTEN antibody [EPR23729-4] (ab260011)
  • Flow Cytometry (Intracellular) - Anti-PTEN antibody [EPR23729-4] (ab260011)
  • Western blot - Anti-PTEN antibody [EPR23729-4] (ab260011)
  • Flow Cytometry (Intracellular) - Anti-PTEN antibody [EPR23729-4] (ab260011)
  • Western blot - Anti-PTEN antibody [EPR23729-4] (ab260011)
  • Immunoprecipitation - Anti-PTEN antibody [EPR23729-4] (ab260011)
  • Anti-PTEN antibody [EPR23729-4] (ab260011)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23729-4] to PTEN
  • Suitable for: Flow Cyt (Intra), IP, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-PTEN antibody [EPR23729-4]
    See all PTEN primary antibodies

  • Description

    Rabbit monoclonal [EPR23729-4] to PTEN

  • Host species

    Rabbit

  • Tested applications

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

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Wild-type HAP1 whole cell lysate; MCF7, RAW 264.7, PC-12, NIH/3T3, Neuro 2a and HeLa whole cell lysates. Flow Cyt (intra): Wild-type HAP1, MCF7 and NIH/3T3 cells. IP: MCF7 and NIH/3T3 whole 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 ab260011 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)
1/60.
IP
1/30.
WB
1/1000. Detects a band of approximately 54 kDa (predicted molecular weight: 47 kDa).
Notes
Flow Cyt (Intra)
1/60.
IP
1/30.
WB
1/1000. Detects a band of approximately 54 kDa (predicted molecular weight: 47 kDa).
Application notes
Is unsuitable for ICC/IF or IHC-P.

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 [EPR23729-4] (ab260011)
    Western blot - Anti-PTEN antibody [EPR23729-4] (ab260011)
    All lanes : Anti-PTEN antibody [EPR23729-4] (ab260011) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : PTEN knockout HAP1 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

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



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

    Exposure time: 59 seconds.

  • Flow Cytometry (Intracellular) - Anti-PTEN antibody [EPR23729-4] (ab260011)
    Flow Cytometry (Intracellular) - Anti-PTEN antibody [EPR23729-4] (ab260011)

    Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized Parental HAP1 (Wild-type control Human chronic myelogenous leukemia near-haploid cell line) (Right) / PTEN KO HAP1 (Left) cells labelling PTEN with ab260011 at 1/600 dilution (0.1ug) (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. 

     

     

  • Immunoprecipitation - Anti-PTEN antibody [EPR23729-4] (ab260011)
    Immunoprecipitation - Anti-PTEN antibody [EPR23729-4] (ab260011)

    PTEN was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast), whole cell lysate with ab260011 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab260011 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: NIH/3T3 (mouse embryonic fibroblast), whole cell lysate 10 ug

    Lane 2: ab260011 IP in NIH/3T3 whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab260011 in NIH/3T3 whole cell lysate

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

    Exposure time: 3 minutes.

     

  • Flow Cytometry (Intracellular) - Anti-PTEN antibody [EPR23729-4] (ab260011)
    Flow Cytometry (Intracellular) - Anti-PTEN antibody [EPR23729-4] (ab260011)

    Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast) cells labelling PTEN with ab260011 at 1/60 dilution (1ug) (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. 

     

     

  • Western blot - Anti-PTEN antibody [EPR23729-4] (ab260011)
    Western blot - Anti-PTEN antibody [EPR23729-4] (ab260011)
    All lanes : Anti-PTEN antibody [EPR23729-4] (ab260011) 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 : Neuro-2a (mouse neuroblastoma neuroblast), whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

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



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

    Exposure times: Lane 1-4: 26 seconds Lane 5: 5.5 seconds.

  • Flow Cytometry (Intracellular) - Anti-PTEN antibody [EPR23729-4] (ab260011)
    Flow Cytometry (Intracellular) - Anti-PTEN antibody [EPR23729-4] (ab260011)

    Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized MCF7 (Human breast adenocarcinoma epithelial cell) cells labelling PTEN with ab260011 at 1/60 dilution (1ug) (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. 

     

     

  • Western blot - Anti-PTEN antibody [EPR23729-4] (ab260011)
    Western blot - Anti-PTEN antibody [EPR23729-4] (ab260011)
    All lanes : Anti-PTEN antibody [EPR23729-4] (ab260011) at 1/1000 dilution

    Lane 1 : MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate
    Lane 2 : MDA-MB-468 (human breast adenocarcinoma epithelial cell), whole cell lysate
    Lane 3 : 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), Peroxidase conjugated (ab97051) at 1/100000 dilution

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



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

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

    Exposure time: 26 seconds.

  • Immunoprecipitation - Anti-PTEN antibody [EPR23729-4] (ab260011)
    Immunoprecipitation - Anti-PTEN antibody [EPR23729-4] (ab260011)

    PTEN was immunoprecipitated from 0.35 mg MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate with ab260011 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab260011 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 10 ug

    Lane 2: ab260011 IP in MCF7 whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab260011 in MCF7 whole cell lysate

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

    Exposure time: 3 minutes.

     

  • Anti-PTEN antibody [EPR23729-4] (ab260011)
    Anti-PTEN antibody [EPR23729-4] (ab260011)

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 (0)

Publishing research using ab260011? Please let us know so that we can cite the reference in this datasheet.

ab260011 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

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