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Cell Biology Cell Cycle Kinases/Phosphatases Phosphatases
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Human PTEN knockout HeLa cell lysate (ab263829)

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Western blot - Human PTEN knockout HeLa cell lysate (ab263829)
  • Western blot - Human PTEN knockout HeLa cell lysate (ab263829)
  • Sanger Sequencing - Human PTEN knockout HeLa cell lysate (ab263829)
  • Sanger Sequencing - Human PTEN knockout HeLa cell lysate (ab263829)
  • Sanger Sequencing - Human PTEN knockout HeLa cell lysate (ab263829)

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Primary
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Anti-PTEN antibody [EPR4408-76] (ab133532)
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Anti-PTEN antibody [EPR9941] (ab154812)

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Overview

  • Product name

    Human PTEN knockout HeLa cell lysate
    See all PTEN kits
  • Product overview


    Knockout cell lysate achieved by CRISPR/Cas9.

  • Parental Cell Line

    HeLa
  • Organism

    Human
  • Mutation description

    Knockout achieved by using CRISPR/Cas9, 11 bp deletion in exon5 and 5 bp insertion in exon5 and Insertion of the selection cassette in exon5.
  • Passage number

    <20
  • Knockout validation

    Sanger Sequencing, Western Blot (WB)
  • Reconstitution notes

    To use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.

    *Usage of SDS sample buffer is not recommended with these lyophilized lysates.

  • Notes

    Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version - found here. Please refer to our lysis protocol for further details on how our lysates are prepared.

    User storage instructions: After reconstitution, store the lysate at -80°C.

    Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
    See here for more information on knockout cell lysates.

    Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
    It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

    This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.

  • Tested applications

    Suitable for: WBmore details

Properties

  • Storage instructions

    Store at -80°C. Please refer to protocols.
  • Components 1 kit
    ab255523 - Human PTEN knockout HeLa cell lysate (Lyophilized) 1 x 100µg
    ab255929 - Human Wild Type HeLa cell lysate (Lyophilized) 1 x 100µg
  • Research areas

    • Cell Biology
    • Cell Cycle
    • Kinases/Phosphatases
    • Phosphatases
    • Signal Transduction
    • Signaling Pathway
    • Lipid Signaling
    • Lipid Phosphatases
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Cancer susceptibility
    • Tumor Suppressors
    • Cancer
    • Cell cycle
    • Kinases/phosphatases
    • Phosphatases
    • Cancer
    • Oncoproteins/suppressors
    • Tumor suppressors
    • PTEN pathway
    • Metabolism
    • Types of disease
    • Diabetes
    • Metabolism
    • Types of disease
    • Obesity
    • Metabolism
    • Types of disease
    • Metabolic disorders
  • Cell type

    epithelial
  • Disease

    Adenocarcinoma
  • Gender

    Female
  • STR Analysis

    Amelogenin X D5S818: 11, 12 D13S317: 12, 13.3 D7S820: 8, 12 D16S539: 9, 10 vWA: 16, 18 TH01: 7 TPOX: 8,12 CSF1PO: 9, 10

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.
  • Target information above from: UniProt accession P60484 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Alternative names

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

Associated products

  • KO cell lines

    • Human PTEN knockout HeLa cell line (ab255419)
  • Related Products

    • Anti-PTEN antibody [EPR4408-76] (ab133532)
    • Anti-PTEN antibody [EPR9941] (ab154812)
    • Anti-PTEN antibody [EPR4408-76] - BSA and Azide free (ab248537)
    • Anti-PTEN antibody [EPR9941] - BSA and Azide free (ab249119)

Applications

Our Abpromise guarantee covers the use of ab263829 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.

Images

  • Western blot - Human PTEN knockout HeLa cell lysate (ab263829)
    Western blot - Human PTEN knockout HeLa cell lysate (ab263829)
    Lane 1: Wild-type HeLa cell lysate (20µg)

    Lane 2: PTEN knockout HeLa cell lysate (20µg)

    Lanes 1- 2: Merged signal (red and green). Green - ab154812 observed at 47 kDa. Red - loading control ab8245 observed at 37 kDa.

    ab154812 Recombinant Anti-PTEN antibody [EPR9941] was shown to specifically 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 and PTEN knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab154812 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 10000 and 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 - Human PTEN knockout HeLa cell lysate (ab263829)
    Western blot - Human PTEN knockout HeLa cell lysate (ab263829)
    Lane 1: Wild-type HeLa cell lysate (20µg)

    Lane 2: PTEN knockout HeLa cell lysate (20µg)

    Lanes 1- 2: Merged signal (red and green). Green - ab133532 observed at 47 kDa. Red - loading control ab130007 observed at 124 kDa.

    ab133532 Recombinant Anti-PTEN antibody [EPR4408-76] was shown to specifically 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 and PTEN knockout samples 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] were incubated overnight at 4°C at 1 in 10000 dilution and 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.
  • Sanger Sequencing - Human PTEN knockout HeLa cell lysate (ab263829)
    Sanger Sequencing - Human PTEN knockout HeLa cell lysate (ab263829)

    Allele-1: 11 bp deletion in exon5

     

  • Sanger Sequencing - Human PTEN knockout HeLa cell lysate (ab263829)
    Sanger Sequencing - Human PTEN knockout HeLa cell lysate (ab263829)

    Allele-2: 5 bp insertion in exon5

     

  • Sanger Sequencing - Human PTEN knockout HeLa cell lysate (ab263829)
    Sanger Sequencing - Human PTEN knockout HeLa cell lysate (ab263829)

    Allele-3: Insertion of the selection cassette in exon5

     

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

    • Datasheet
    • SDS
  • References (0)

    Publishing research using ab263829? Please let us know so that we can cite the reference in this datasheet.

    ab263829 has not yet been referenced specifically in any publications.

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