PTN KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout.
Images
Key facts
- Cell type
- U-87 MG
- Species or organism
- Human
- Tissue
- Brain
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout.
Recommended products
PTN KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout.
Key facts
- Cell type
- U-87 MG
- Species or organism
- Human
- Tissue
- Brain
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout.
- Disease
- Glioblastoma
- Concentration
Properties
- Gene name
- PTN
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing, Western blot
Cell culture
- Biosafety level
- EU: 1 US: 1
- Viability
- ~ 80%
- Adherent/suspension
- Adherent
- Gender
- Male
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
- EMEM + 10% FBS
- Cryopreservation medium
- Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.
Recommended control: Human wild-type U-87 MG cell line (ab278079). 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: EMEM + 10% FBS
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. Based on cell count, seed cells in an appropriate cell culture flask at a density of 2x10E4 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules.
4. Incubate the culture at 37°C 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 2x10E4 cells/cm2 is recommended.
- 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.
We will provide viable cells that proliferate on revival.
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Pleiotrophin also known as PTN Ptn or $Ptn is a heparin-binding growth factor with a molecular mass of approximately 18 kDa. It is expressed in various tissues notably in the brain during embryonic development and also in adulthood. PTN plays a fundamental role in cellular proliferation differentiation and survival. As a secreted protein it interacts with cell surface receptors to trigger intracellular signaling pathways.
- Biological function summary
Pleiotrophin functions as a potent mitogen promoting cell growth and angiogenesis. It does not operate as part of a larger protein complex but acts independently to influence cellular behavior. Its interaction with receptor protein tyrosine phosphatase (RPTP) modulates the tyrosine phosphorylation state of other proteins affecting downstream signaling. This interaction facilitates communication between cells and their environments influencing development and tissue maintenance.
- Pathways
This protein significantly impacts the PI3K/Akt and MAPK/ERK pathways key regulators of cell survival and proliferation. PTN often collaborates with proteins like Midkine (MK) due to their functional similarities and overlapping receptor targets. These pathways help regulate cellular responses to environmental cues contributing to processes such as tissue repair and regeneration.
- Associated diseases and disorders
The overexpression of pleiotrophin has been linked to the development of cancers such as breast cancer and glioblastoma. PTN's interaction with oncogenic pathways often involving proteins like N-cadherin enhances its relevance in tumor progression and metastasis. Additionally alterations in PTN expression or activity can influence neural disorders highlighting its importance in both oncological and neurological contexts.
Product promise
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
3 product images
Western blot - Human PTN knockout U-87 MG cell line (ab288713)
Anti-PTN antibody [EP3040(2)] ab133517 was shown to react with PTN in wild-type U87mg cells in Western blot with loss of signal observed in PTN knockout cell line ab288713. Wild-type U87mg and PTN knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with Anti-PTN antibody [EP3040(2)] ab133517 overnight at 4 °C at a 1/500 dilution. Blots were incubated with secondary antibodies at 0.2ug/mL before imaging.
These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.All lanes: Western blot - Anti-PTN antibody [EP3040(2)] (Anti-PTN antibody [EP3040(2)] ab133517) at 1/500 dilution
Lane 1: Wild-type U-87 MG lysate at 25 µg
Lane 2: PTN knock-out U87mg lysate at 25 µg
Western blot - Human PTN knockout U-87 MG cell line (ab288713)
Anti-PTN antibody [EPR3041] ab79411 was shown to react with PTN in wild-type U87mg cells in Western blot with loss of signal observed in PTN knockout cell line ab288713. Wild-type U87mg and PTN knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with Anti-PTN antibody [EPR3041] ab79411 overnight at 4 °C at a 1/500 dilution. Blots were incubated with secondary antibodies at 0.2ug/mL before imaging.
These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
All lanes: Western blot - Anti-PTN antibody [EPR3041] (Anti-PTN antibody [EPR3041] ab79411) at 1/500 dilution
Lanes 1 - 2: Western blot - Human PTN knockout U-87 MG cell line (ab288713)
Lane 1: Wild-type U87mg lysate at 20 µg
Lane 2: PTN Knockout U87mg lysate at 20 µg
Observed band size: 19 kDa
Sanger Sequencing - Human PTN knockout U-87 MG cell line (ab288713)
Functional Homozygote. 38 bp and 37 bp deletions
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