JavaScript is disabled in your browser. Please enable JavaScript to view this website.
AB257810

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

Be the first to review this product! Submit a review

|

(0 Publication)

ZYX KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 2.
4 Images
Western blot - Human ZYX (Zyxin) knockout HEK-293T cell lysate (AB257810)
  • WB

Lab

Western blot - Human ZYX (Zyxin) knockout HEK-293T cell lysate (AB257810)

Lane 1 : Wild-type HEK-293T cell lysate 20 μg
Lane 2 : ZYX knockout HEK-293T cell lysate 20 μg
Lane 3 : HeLa cell lysate 20 μg
Lane 4 : Daudi cell lysate 20 μg
Lanes 1 - 4 : Merged signal (red and green). Green - ab50391 observed at 75 kDa. Red - loading control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37kDa.
ab50391 was shown to react with ZYX in wild-type HEK-293T cells in Western blot with loss of signal observed in ZYX knockout cell line ab266504 (ZYX knockout cell lysate ab257810). Wild-type HEK-293T and ZYX knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab50391 and ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-Zyxin antibody [ZOL301] (<a href='/en-us/products/primary-antibodies/zyxin-antibody-zol301-ab50391'>ab50391</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

ZYX knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human ZYX (Zyxin) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-zyx-zyxin-knockout-hek-293t-cell-line-ab266504'>ab266504</a>)

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

Western blot - Human ZYX (Zyxin) knockout HEK-293T cell lysate (AB257810)
  • WB

Lab

Western blot - Human ZYX (Zyxin) knockout HEK-293T cell lysate (AB257810)

Lane 1 : Wild-type HEK-293T cell lysate 20 μg

Lane 2 : ZYX knockout HEK-293T cell lysate 20 μg

Lane 3 : HeLa cell lysate 20 μg

Lane 4 : Daudi cell lysate 20 μg

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 70 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in ZYX knockout cell line ab266504 (knockout cell lysate ab257810). The band observed in the knockout lysate lane below 70 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 knockout HEK-293T cell lysate at 20 µg

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

Western blot - Human ZYX (Zyxin) knockout HEK-293T cell lysate (AB257810)
  • WB

Lab

Western blot - Human ZYX (Zyxin) knockout HEK-293T cell lysate (AB257810)

Lane 1 : Wild-type HEK-293T cell lysate 20 μg
Lane 2 : ZYX knockout HEK-293T cell lysate 20 μg
Lane 3 : HeLa cell lysate 20 μg
Lane 4 : Daudi cell lysate 20 μg
Lanes 1 - 4 : Merged signal (red and green). Green - ab229757 observed at 75 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab229757 was shown to react with ZYX in wild-type HEK-293T cells in Western blot with loss of signal observed in ZYX knockout cell line ab266504 (ZYX knockout cell lysate ab257810). Wild-type HEK-293T and ZYX knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab229757 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

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 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human ZYX (Zyxin) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-zyx-zyxin-knockout-hek-293t-cell-line-ab266504'>ab266504</a>)

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 lysate (AB257810)
  • Sanger seq

Unknown

Sanger Sequencing - Human ZYX (Zyxin) knockout HEK-293T cell lysate (AB257810)

Homozygous : Insertion of the selection cassette in exon 2

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 2.

style
left-column

Complementary Products

Primary Antibodies

AB109316

Anti-Zyxin antibody [EPR4302]

BOND RX™ Validated|RabMAb|Recombinant

Flow Cytometry (Intracellular) - Anti-Zyxin antibody [EPR4302] (AB109316)

  • 0
  • 0
Primary Antibodies

AB60933

Anti-Drebrin antibody

KO Validated

Immunoprecipitation - Anti-Drebrin antibody (AB60933)

  • 5
  • 6
Primary Antibodies

AB71333

Anti-Talin 1 antibody

BOND RX™ Validated|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Talin 1 antibody (AB71333)

  • 4
  • 4
Cell Lines & Lysates

AB266503

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

Advanced Validation

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

  • 0
  • 0
style
left-column

Reactivity data

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

Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.

style
left-column

What's included?

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab257810-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human ZYX knockout HEK293T cell lysate", "number":"AB257810-CMP01", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type HEK293T cell lysate", "number":"AB257810-CMP02", "productcode":"" } ] } } }
style
left-column

Properties and storage information

Gene name
ZYX
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
style
left-column

Supplementary information

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.
style
left-column

Quality control

STR analysis

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

style
left-column

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

style
left-column

Product protocols

style
left-column
style
left-column
style
right-column

Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

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