ZYX KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 19 bp deletion in exon 2 and 1 bp deletion in exon 2.
Images
Key facts
- Cell type
- HEK-293T
- Species or organism
- Human
- Tissue
- Kidney
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 19 bp deletion in exon 2 and 1 bp deletion in exon 2
Alternative names
ESP 2, HED 2, ZYX protein, ZYX_HUMAN, Zyxin, Zyxin-2
Recommended products
ZYX KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 19 bp deletion in exon 2 and 1 bp deletion in exon 2.
Key facts
- Cell type
- HEK-293T
- Species or organism
- Human
- Tissue
- Kidney
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 19 bp deletion in exon 2 and 1 bp deletion in exon 2
- Concentration
Properties
- Gene name
- ZYX
- 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
Cell culture
- Biosafety level
- EU: 2 US: 2
- Adherent/suspension
- Adherent
- Gender
- Female
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.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
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
Zyxin also known as ZYX is a mechanical regulator associated with cell adhesion and signaling. This protein has a molecular mass of approximately 61 kDa and shows expression in various cell types including those in the heart liver and lungs. As a component of focal adhesions zyxin plays a role in cytoskeletal dynamics by interacting with actin filaments and facilitating the formation of stress fibers.
- Biological function summary
The protein has important roles in cellular processes such as migration and morphology. Zyxin often forms part of a larger protein complex that includes VASP and α-actinin which contribute to cell scaffolding and structural integrity. It serves as an organizer for cytoskeletal proteins ensuring proper cell movement and adaptability to cellular stress. Its actin-binding capacity enables it to regulate structural changes necessary for cellular adaptation to environmental stimuli.
- Pathways
This protein is essential in the regulation of actin cytoskeleton associated pathways and adherens junction signaling. Zyxin interacts with Ena/VASP proteins which are significant in actin filament elongation helping regulate dynamic changes in cell shape and motility. Its relationship with LIM domain-containing proteins positions zyxin within pathways related to cell proliferation and signal transduction.
- Associated diseases and disorders
Zyxin has been implicated in cancer progression especially in metastasis where altered cell adhesion and movement play a role. It also relates to cardiovascular diseases due to its involvement in mechanotransduction pathways critical to heart muscle function. In cancer zyxin influences the activity of YAP/TAZ transcription regulators affecting tumor cell behavior. In cardiovascular scenarios the protein interacts with paxillin to mediate responses to mechanical load changes impacting heart disease progression.
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4 product images
Western blot - Human ZYX (Zyxin) knockout HEK-293T cell line (ab266503)
False colour image of Western blot: Anti-Zyxin antibody [EPR4302] staining at 1/20000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution,shown in red. In Western blot,Anti-Zyxin antibody [EPR4302] ab109316 was shown to bind specifically to Zyxin. A band was observed at 75 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in ZYX knockout cell line ab266503 (knockout cell lysate Human ZYX (Zyxin) knockout HEK-293T cell lysate ab257809). The band observed in the knockout lysate lane below 75 kDa is likely to represent a truncated form of Zyxin. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and ZYX knockout HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution 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 (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-Zyxin antibody [EPR4302] (Anti-Zyxin antibody [EPR4302] ab109316) at 1/20000 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: ZYX CRISPR-Cas9 edited HEK-293T cell lysate at 20 µg
Lane 2: Western blot - Human ZYX (Zyxin) knockout HEK-293T cell line (ab266503)
Lane 3: HeLa cell lysate at 20 µg
Lane 4: Daudi cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 61 kDa
Observed band size: 75 kDa
Western blot - Human ZYX (Zyxin) knockout HEK-293T cell line (ab266503)
False colour image of Western blot: Anti-Zyxin antibody - N-terminal staining at 1/1000 dilution,shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution,shown in red. In Western blot.Anti-Zyxin antibody - N-terminal ab229757 was shown to bind specifically to Zyxin. A band was observed at 75 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in ZYX knockout cell line ab266503 (knockout cell lysate Human ZYX (Zyxin) knockout HEK-293T cell lysate ab257809). The band observed in the knockout lysate lane below 75 kDa is likely to represent a truncated form of Zyxin. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image,wild-type and ZYX knockout HEK-293T cell lysates were analysed. First,samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 %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 (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-Zyxin antibody - N-terminal (Anti-Zyxin antibody - N-terminal ab229757) at 1/1000 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: ZYX CRISPR-Cas9 edited HEK-293T cell lysate at 20 µg
Lane 2: Western blot - Human ZYX (Zyxin) knockout HEK-293T cell line (ab266503)
Lane 3: HeLa cell lysate at 20 µg
Lane 4: Daudi cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 61 kDa
Observed band size: 75 kDa
Sanger Sequencing - Human ZYX (Zyxin) knockout HEK-293T cell line (ab266503)
Allele-1: 19 bp deletion in exon2
Sanger Sequencing - Human ZYX (Zyxin) knockout HEK-293T cell line (ab266503)
Allele-2: 1 bp deletion in exon 2.
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