Human PTEN knockout HeLa cell line
- Advanced Validation
- What is this?
Be the first to review this product! Submit a review
|
(0 Publication)
PTEN KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. 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 5. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View Alternative Names
10q23del, BZS, DEC, GLM2, MGC11227, MHAM, MMAC1, MMAC1 phosphatase and tensin homolog deleted on chromosome 10, Mutated in multiple advanced cancers 1, PTEN1, PTEN_HUMAN, Phosphatase and Tensin Homolog, Phosphatase and tensin like protein, Phosphatidylinositol 3,4,5-trisphosphate 3-phosphatase and dual-specificity protein phosphatase PTEN
- WB
Lab
Western blot - Human PTEN knockout HeLa cell line (AB255419)
Lanes 1- 2 : Merged signal (red and green). Green - ab133532 observed at 47 kDa. Red - Anti-Vinculin antibody [VIN-54] observed at 124 kDa.
ab133532 was shown to 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 HeLa and PTEN knockout HeLa cell lysates 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] overnight at 4° at a 1 in 10000 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.
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 (ab255419)
Predicted band size: 47 kDa
Observed band size: 47 kDa
false
- WB
Lab
Western blot - Human PTEN knockout HeLa cell line (AB255419)
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 (ab255419)
Lane 3:
HAP1 cell lysate at 20 µg
Predicted band size: 47 kDa
Observed band size: 47 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human PTEN knockout HeLa cell line (AB255419)
Allele-3 : Insertion of the selection cassette in exon 5.
- Sanger seq
Unknown
Sanger Sequencing - Human PTEN knockout HeLa cell line (AB255419)
Allele-2 : 5 bp insertion in exon 5.
- Sanger seq
Unknown
Sanger Sequencing - Human PTEN knockout HeLa cell line (AB255419)
Allele-1 : 11 bp deletion in exon 5.
Reactivity data
Product details
We will provide viable cells that proliferate on revival.
Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Handling procedures
Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for 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-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x 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-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. 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 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
Cell Lines & Lysates
AB261755
Human IDE (Insulin degrading enzyme) knockout HeLa cell line
Advanced Validation
- 0
- 0
Primary Antibodies
AB32199
20ul selling size|RabMAb|Recombinant|KO Validated|Trial Size
![Western blot - Anti-PTEN antibody [Y184] (AB32199)](https://content.abcam.com/products/images/pten-antibody-y184-ab32199--western-blot-img153032.jpg)
- 5
- 15
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com