Human ZYX (Zyxin) knockout HEK-293T cell line
- Advanced Validation
- What is this?
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ZYX KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 19 bp deletion in exon 2 and 1 bp deletion in exon 2. 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
ESP 2, HED 2, ZYX protein, ZYX_HUMAN, Zyxin, Zyxin-2
- WB
Lab
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] (ab8245) loading control staining at 1/20000 dilution,shown in red. In Western blot,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 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-Zyxin antibody [EPR4302] (<a href='/en-us/products/primary-antibodies/zyxin-antibody-epr4302-ab109316'>ab109316</a>) 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
Predicted band size: 61 kDa
Observed band size: 75 kDa
false
- WB
Lab
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] (ab8245) loading control staining at 1/20000 dilution,shown in red. In Western blot.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 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-Zyxin antibody - N-terminal (<a href='/en-us/products/primary-antibodies/zyxin-antibody-n-terminal-ab229757'>ab229757</a>) 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
Predicted band size: 61 kDa
Observed band size: 75 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human ZYX (Zyxin) knockout HEK-293T cell line (AB266503)
Allele-2 : 1 bp deletion in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human ZYX (Zyxin) knockout HEK-293T cell line (AB266503)
Allele-1 : 19 bp deletion in exon2
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 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.
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
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.
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
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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