TLN1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout.
Images
Key facts
- Cell type
- PC-3
- Species or organism
- Human
- Tissue
- Prostate
- Form
- Liquid
- Knockout validation
- Western blot
- Mutation description
- Knockout.
Alternative names
ILWEQ, TLN, TLN1_HUMAN, Talin, Talin-1
Recommended products
TLN1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout.
Key facts
- Cell type
- PC-3
- Species or organism
- Human
- Tissue
- Prostate
- Form
- Liquid
- Knockout validation
- Western blot
- Mutation description
- Knockout.
- Disease
- Adenocarcinoma
- Concentration
Properties
- Gene name
- TLN1
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Western blot
Cell culture
- Biosafety level
- EU: 1 US: 1
- 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
- F-12K + 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
Recommended control: Human wild-type PC-3 cell line (ab290718). 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: F-12K + 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 2x104 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 2x104 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
Talin 1 (TLN1) is a cytoskeletal protein that mechanically links integrins to actin filaments in cells. It has a molecular weight of 270 kDa and is expressed in many tissues including muscle brain and kidney. Talin 1 contains a head domain and a tail domain which bind to integrins and actin respectively. By establishing these connections Talin 1 plays a role in cell adhesion spreading and migration. This protein also interacts with other adhesion-associated proteins reinforcing its structural function inside cells.
- Biological function summary
Talin 1 is involved in facilitating integrin activation and focal adhesion formation. It is a component of a larger complex that includes proteins like vinculin and paxillin. These complexes are responsible for transmitting mechanical signals from the extracellular matrix into the intracellular environment initiating cellular responses like growth and differentiation. Talin 1 also modulates actin dynamics which contributes to changes in cell shape and motility. Its role in these processes highlights its significance in maintaining cellular architecture.
- Pathways
Talin 1 participates in the integrin-mediated signaling pathway and the Rho GTPase signaling pathway. In the integrin pathway Talin 1 interacts with integrin beta subunits leading to integrin clustering and increased focal adhesion strength. In the Rho GTPase pathway Talin 1 influences actin cytoskeleton rearrangements through its interactions with proteins like RhoA and Rac1. These pathways are critical for processes such as cell proliferation and migration which are essential for proper tissue function and development.
- Associated diseases and disorders
Talin 1 has an association with cancer and cardiovascular diseases. Changes in Talin 1 expression and localization can contribute to altered cell adhesion and motility promoting cancer cell invasion and metastasis. Talin 1 has a relationship with the protein FAK (Focal Adhesion Kinase) in cancer progression. In cardiovascular diseases Talin 1 dysregulation affects cardiomyocyte adhesion and survival and it interacts with proteins such as vinculin to sustain cardiac tissue integrity. These connections highlight Talin 1's role in disease pathology.
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2 product images
Western blot - Human TLN1 knockout PC-3 cell line (ab290495)
Western blot: Anti-TLN1 antibody (Anti-Talin 1 antibody ab71333) staining at 1 ug/ml, shown in green; Mouse anti-CANX [CANX/1543] (Anti-Calnexin antibody [CANX/1543] ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-Talin 1 antibody ab71333 was shown to bind specifically to TLN1. A band was observed at 270 kDa in wild-type PC-3 cell lysates with no signal observed at this size in TLN1 knockout cell line. To generate this image, wild-type and TLN1 knockout PC-3 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-Talin 1 antibody (Anti-Talin 1 antibody ab71333) at 1 µg/mL
Lane 1: Wild-type PC-3 cell lysate at 20 µg
Lane 2: TLN1 knockout PC-3 cell lysate at 20 µg
Lane 3: TLN1 knockout PC-3 cell lysate at 10 µg
Lane 4: HeLa cell lysate at 20 µg
Lane 5: SK-OV-3 cell lysate at 20 µg
SecondaryAll lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 270 kDa
Next Generation Sequencing - Human TLN1 knockout PC-3 cell line (ab290495)
104 bp deletion after Cys762 (allele 1) and 58 bp deletion after Lys744 (allele 2) of the WT protein
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