Human MVP knockout HeLa cell line (ab264817)

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Overview

  • Product name

    Human MVP knockout HeLa cell line

  • Parental Cell Line

    HeLa

  • Organism

    Human

  • Mutation description

    Knockout achieved by using CRISPR/Cas9, 17 bp deletion in exon 6 and 1 bp deletion in exon 6

  • Passage number

    <20

  • Knockout validation

    Sanger Sequencing, Western Blot (WB)

  • Biosafety level

    2

  • General notes

    Recommended control: Human Wild Type HeLa cell line (ab255448)

    Please note a wild type cell line is not automatically included with a KO cell line order, if required please add recommended wild type cell line at no additional cost using the code WILDTYPE-TMTK1

    Cryopreservation cell medium: Cell Freezing Medium-DMSO Serum free media, supplemented with 10% (v/v) DMSO

    Handling procedure: Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C will result in loss of viability.
    1. Thaw the vial in 37°C water bath approximately 1-2 minutes.
    2. Transfer the cell suspension to a 15 mL conical tube with pre-warmed 5 mL complete medium DMEM+10% FBS, spin 125×g for approximately 5 minutes at room temperature.
    3. Resuspend the cell pellet with 1 mL pre-warmed complete medium DMEM+10% FBS and dispense into a 25 cm2 culture flask containing 10 mL pre-warmed complete complete medium DMEM+10% FBS.
    4. Incubate the culture at 37°C incubator with 5% CO2.
    5. A subcultivation ratio of 1:4-1:6 is recommended. Cells should be passaged when cells grow splitting at 80-90% confluence.

    Click here to view the Mammalian cell tissue culture protocol

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

Properties

  • Number of cells

    1 x 106 cells/vial, 1 mL

  • Viability

    ~90%

  • Adherent /Suspension

    Adherent

  • Tissue

    Cervix

  • 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

  • Antibiotic resistance

    Puromycin 1.00µg/ml

  • Mycoplasma free

    Yes

  • Storage instructions

    Shipped on Dry Ice. Store in liquid nitrogen.

  • Storage buffer

    Constituents: 8.7% DMSO, 2% Cellulose, methyl ether

  • Research areas

Target

  • Function

    Required for normal vault structure. Vaults are multi-subunit structures that may act as scaffolds for proteins involved in signal transduction. Vaults may also play a role in nucleo-cytoplasmic transport. Down-regulates INFG-mediated STAT1 signaling and subsequent activation of JAK. Down-regulates SRC activity and signaling through MAP kinases.

  • Tissue specificity

    Present in most normal tissues. Higher expression observed in epithelial cells with secretory and excretory functions, as well as in cells chronically exposed to xenobiotics, such as bronchial cells and cells lining the intestine. Overexpressed in many multidrug-resistant cancer cells.

  • Sequence similarities

    Contains 9 MVP (vault) repeats.

  • Domain

    MVP 3 mediates interaction with PTEN.
    MVP 4 mediates interaction with PARP4.

  • Post-translational
    modifications

    Phosphorylated on Tyr residues after EGF stimulation.
    Dephosphorylated by PTPN11.

  • Cellular localization

    Cytoplasm. Nucleus > nuclear pore complex. 5% found in the nuclear pore complex. Translocates from the nucleus to the cytoplasm upon EGF treatment.
  • Information by UniProt

Images

  • Western blot - Human MVP knockout HeLa cell line (ab264817)
    Western blot - Human MVP knockout HeLa cell line (ab264817)
    All lanes : Anti-MVP antibody [EPR13226(B)] (ab177145) at 1/1000 dilution

    Lane 1 : Wild-type HeLa lysate
    Lane 2 : MVP knockout HeLa lysate
    Lane 3 : A549 lysate
    Lane 4 : MOLT-4 lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.


    Lanes 1- 4: Merged signal (red and green). Green - ab177145 observed at 99 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

    ab177145 was shown to react with MVP in wild-type HeLa cells in Western blot. Loss of signal was observed when knockout sample ab257544 was used. Wild-type HeLa and MVP knockout HeLa whole cell lysates were subjected to SDS-PAGE. ab177145 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 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 - Human MVP knockout HeLa cell line (ab264817)
    Western blot - Human MVP knockout HeLa cell line (ab264817)
    All lanes : Anti-MVP antibody [EPR13227(B)] (ab175239) at 1/2000 dilution

    Lane 1 : Wild-type HeLa lysate
    Lane 2 : MVP knockout HeLa lysate
    Lane 3 : A549 lysate
    Lane 4 : MOLT-4 lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.


    Lanes 1- 4: Merged signal (red and green). Green - ab175239 observed at 110 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

    ab175239 was shown to react with MVP in wild-type HeLa cells in Western blot. Loss of signal was observed when knockout sample ab257544 was used. Wild-type HeLa and MVP knockout HeLa whole cell lysates were subjected to SDS-PAGE. ab175239 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 2000 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.

  • Sanger Sequencing - Human MVP knockout HeLa cell line (ab264817)
    Sanger Sequencing - Human MVP knockout HeLa cell line (ab264817)
    Allel-1: 17 bp deletion in exon 6.
  • Sanger Sequencing - Human MVP knockout HeLa cell line (ab264817)
    Sanger Sequencing - Human MVP knockout HeLa cell line (ab264817)
    Allel-2: 1 bp deletion in exon 6.

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 ab264817? Please let us know so that we can cite the reference in this datasheet.

ab264817 has not yet been referenced specifically in any publications.

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