AB266503

人ZYX (Zyxin) knockout HEK-293T cell line

Human ZYX (Zyxin) knockout HEK-293T cell line

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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 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

查看别名

ESP 2, HED 2, ZYX protein, ZYX_HUMAN, Zyxin, Zyxin-2

4 Images

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='/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

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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='/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 Sequencing - Human ZYX (Zyxin) knockout HEK-293T cell line (AB266503)

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

关键信息

细胞类型

HEK-293T

种属

Human

组织

Kidney

形式

Liquid

form

敲除验证

Sanger Sequencing,Western blot

突变描述

Knockout achieved by using CRISPR/Cas9, 19 bp deletion in exon 2 and 1 bp deletion in exon 2

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反应性数据

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

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产品详情

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.

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规格

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性能和储存信息

基因名称

ZYX

基因编辑类型

Knockout

基因编辑方法

CRISPR technology

敲除验证

Sanger Sequencing, Western blot

运输条件

Dry Ice

处理步骤

初始处理指南

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.

传代培养指南

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.

培养基

DMEM (High Glucose) + 10% FBS

低温储藏试剂

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

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补充信息

This supplementary information is collated from multiple sources and compiled automatically.

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.

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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质量控制

STR 分析

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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细胞培养

生物安全等级

EU: 2 US: 2

贴壁/悬浮

Adherent

性别

Female

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产品实验方案

Visit the General protocols
Visit the Troubleshooting

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靶点信息

See full target information ZYX

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Abcam Product Promise

我们致力于为您的研究提供高质量的试剂，为您科研的每一步提供支持。若我们的产品未能达到预期性能，我们向您提供 Abcam Product Promise 保障。

详情请参阅我们的条款与条件。

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

人ZYX (Zyxin) knockout HEK-293T cell line

Human ZYX (Zyxin) knockout HEK-293T cell line

Western blot - Human ZYX (Zyxin) knockout HEK-293T cell line (AB266503)

Western blot - Human ZYX (Zyxin) knockout HEK-293T cell line (AB266503)

Sanger Sequencing - Human ZYX (Zyxin) knockout HEK-293T cell line (AB266503)

Sanger Sequencing - Human ZYX (Zyxin) knockout HEK-293T cell line (AB266503)

关键信息

细胞类型

种属

组织

形式

敲除验证

突变描述

反应性数据

产品详情

规格

性能和储存信息

基因名称

基因编辑类型

基因编辑方法

敲除验证

运输条件

推荐的短期储存条件

推荐的长期储存条件

处理步骤

初始处理指南

传代培养指南

培养基

低温储藏试剂

补充信息

Biological function summary

Pathways

质量控制

STR 分析

细胞培养

生物安全等级

贴壁/悬浮

性别

产品实验方案

靶点信息

搭配使用产品

Primary Antibodies

Cell Lines & Lysates

Primary Antibodies

Primary Antibodies

Abcam Product Promise