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AB263829

Human PTEN knockout HeLa cell lysate

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PTEN KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. 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.
6 Images
Western blot - Human PTEN knockout HeLa cell lysate (AB263829)
  • WB

Lab

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.

All lanes:

Western blot - Anti-PTEN antibody [EPR4408-76] (<a href='/en-us/products/primary-antibodies/pten-antibody-epr4408-76-ab133532'>ab133532</a>) at 1/10000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

PTEN knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human PTEN knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-pten-knockout-hela-cell-line-ab255419'>ab255419</a>)

Predicted band size: 47 kDa

Observed band size: 47 kDa

false

Western blot - Human PTEN knockout HeLa cell lysate (AB263829)
  • WB

Lab

Western blot - Human PTEN knockout HeLa cell lysate (AB263829)

Lane 1 : Wild-type HeLa cell lysate 20 ug
Lane 2 : PTEN knockout HeLa cell lysate 20 ug
Lane 3 : HAP1 cell lysate 20 ug

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 bands observed in PTEN knockout cell line ab255419 (PTEN knockout cell lysate ab263829) below 47 kDa may represent truncated forms and cleaved fragments. This has not been investigated further. HeLa wild-type and PTEN knockout 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.

All lanes:

Western blot - Anti-PTEN antibody [EPR4408-76] (<a href='/en-us/products/primary-antibodies/pten-antibody-epr4408-76-ab133532'>ab133532</a>) at 1/10000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

PTEN knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human PTEN knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-pten-knockout-hela-cell-line-ab255419'>ab255419</a>)

Lane 3:

HAP1 cell lysate at 20 µg

Predicted band size: 47 kDa

Observed band size: 47 kDa

false

Western blot - Human PTEN knockout HeLa cell lysate (AB263829)
  • WB

Lab

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.

All lanes:

Western blot - Anti-PTEN antibody [EPR9941] (<a href='/en-us/products/primary-antibodies/pten-antibody-epr9941-ab154812'>ab154812</a>) at 1/10000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

PTEN knockout HeLa cell lysate at 20 µg

Predicted band size: 47 kDa

Observed band size: 47 kDa

false

Sanger Sequencing - Human PTEN knockout HeLa cell lysate (AB263829)
  • Sanger seq

Unknown

Sanger Sequencing - Human PTEN knockout HeLa cell lysate (AB263829)

Allele-3 : Insertion of the selection cassette in exon5

Sanger Sequencing - Human PTEN knockout HeLa cell lysate (AB263829)
  • Sanger seq

Unknown

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 seq

Unknown

Sanger Sequencing - Human PTEN knockout HeLa cell lysate (AB263829)

Allele-1 : 11 bp deletion in exon5

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing,Western blot

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.

Disease

Adenocarcinoma

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Primary Antibodies

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Recombinant|Trial Size

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Reactivity data

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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
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.

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. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

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.

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What's included?

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Properties and storage information

Gene name
PTEN
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Quality control - STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

The PTEN protein also known as phosphatase and tensin homolog is a phosphatase enzyme with a molecular mass of approximately 47 kDa. It acts mechanically by removing phosphate groups from phosphatidylinositol (345)-trisphosphate (PIP3) converting it to phosphatidylinositol (45)-bisphosphate. PTEN is ubiquitously expressed in various tissues with pronounced presence in the brain lung kidney and testis. This enzyme is an important regulator of cellular functions through its impact on signaling pathways.
Biological function summary

PTEN plays important roles in cellular processes like apoptosis cell proliferation and migration. It negatively regulates the PI3K/AKT signaling pathway critical for cell survival and growth. PTEN is not part of a larger protein complex but interacts with various other proteins modulating its activity. It maintains cellular homeostasis by balancing growth-promoting signals with cell cycle arrest and apoptotic pathways.

Pathways

PTEN is an important component in the PI3K/AKT and mTOR signaling pathways. These pathways regulate cell growth metabolism and survival and are interlinked with insulin signaling and cancer progression. PTEN's function directly interacts with proteins such as AKT and mTOR serving as a checkpoint that ensures controlled cellular proliferation. This positions PTEN as a tumor suppressor inhibiting uncontrolled cell growth via modulation of these pathways.

PTEN mutations or deletions are strongly associated with various types of cancers including breast and prostate cancer. PTEN interaction with the PI3K/AKT pathway influences cancer development often through loss-of-function mutations leading to unrestrained cellular growth. Beyond cancer PTEN mutations also relate to neurological disorders like Autism Spectrum Disorder where it affects signaling pathways involving proteins like mTOR. Understanding PTEN's role aids in unravelling the mechanistic underpinnings of these diseases paving the way for targeted therapeutic interventions.
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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Product promise

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For full details, please see our Terms & Conditions

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