Overview
Product name
Human PTEN knockout HeLa cell lineSee all PTEN lysatesParental Cell Line
HeLaOrganism
HumanMutation description
Knockout achieved by using CRISPR/Cas9, 11 bp deletion in exon 5 and 5 bp insertion in exon 5 and Insertion of the selection cassette in exon 5Passage number
<>Knockout validation
Sanger Sequencing, Western Blot (WB)Tested applications
Suitable for:WBmore detailsBiosafety level
2General notes
Recommended control: Human wild-type HeLa cell line (ab255928). Please note a wild-type cell line is not automatically included with a knockout 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, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Culture medium:DMEM (High Glucose) + 10% FBS
Initial handling guidelines:Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80ºC. Storage at -80ºC may result in loss of viability.
1. Thaw the vial in 37ºC water bath approximately 1-2 minutes.2. Transfer the cell suspension (0.8 ml) to a 15 ml/50 ml conical sterile polypropylene centrifuge tube containing 8.4 ml pre-warmedculture medium, wash vial with an additional 0.8 mlculture medium(total volume 10 ml) to collect remaining cells, and centrifuge at 201x g(rcf) for 5 minutes at room temperature. 10 ml represents minimum recommended dilution. 20 ml represents maximum recommended dilution.3. Resuspend the cell pellet in 5 ml pre-warmedculture mediumand count using a haemocytometer (Click here to view haemocytometer protocol) or alternative cell counting method. Based on cell count, seed cells in an appropriate cell culture flask at a density of 2x104cells/cm2. This should allow for confluency within 48 hours. Seeding density is given as a guide only and should be scaled to align with individual lab schedules.4. Incubate the culture at 37ºC incubator with 5% CO2. Cultures should be monitored daily.
Subculture guidelines:
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104cells/cm2is recommended for confluency (80-90% confluence) within 48 hours.
- A partial media change 24 hours prior to subculture may be helpful to encourage growth, if required.
- Cells should be passaged when they have achieved 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, 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.
Properties
Number of cells
1 x 106 cells/vial, 1 mLViability
~90%Adherent /Suspension
AdherentTissue
CervixCell type
epithelialDisease
AdenocarcinomaGender
FemaleSTR Analysis
Amelogenin XD5S818: 11, 12D13S317: 12, 13.3D7S820: 8, 12D16S539: 9, 10vWA: 16, 18TH01: 7TPOX: 8,12CSF1PO: 9, 10Mycoplasma free
YesStorage instructions
Shipped on Dry Ice. Store in liquid nitrogen.Storage buffer
Constituents: 8.7% DMSO, 2% Cellulose, methyl etherResearch 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
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 1Lhermitte-Duclos diseaseBannayan-Riley-Ruvalcaba syndromeSquamous cell carcinoma of the head and neckEndometrial cancerPTEN 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 2VACTERL association with hydrocephalusProstate cancerMacrocephaly/autism syndromeA 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-translationalmodifications
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
