Human ZYX (Zyxin) knockout HEK-293T cell lysate (ab257809)
製品の概要
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製品名
Human ZYX (Zyxin) knockout HEK-293T cell lysate
Zyxin キット 製品一覧 -
製品の概要
Knockout cell lysate achieved by CRISPR/Cas9.
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Parental Cell Line
HEK293T -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, 19 bp deletion in exon2 and 1 bp deletion in exon2. -
Passage number
<20 -
Knockout validation
Sanger Sequencing, Western Blot (WB) -
Reconstitution notes
To use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT. *Usage of SDS sample buffer is not recommended with these lyophilized lysates. -
特記事項
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 - found here. 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.
Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
See here for more information on knockout cell lysates.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.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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アプリケーション
適用あり: WBmore details
製品の特性
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保存方法
Store at -80°C. Please refer to protocols. -
内容 1 kit ab255553 - Human wild-type HEK293T cell lysate 1 x 100µg ab263592 - Human ZYX knockout HEK293T cell lysate 1 x 100µg -
研究分野
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Cell type
epithelial -
Gender
Female -
STR Analysis
Amelogenin X D5S818: 8, 9 D13S317: 12, 14 D7S820: 11 D16S539: 9, 13 vWA: 16, 19 TH01: 7, 9.3 TPOX: 11 CSF1PO: 11, 12
ターゲット情報
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機能
Adhesion plaque protein. Binds alpha-actinin and the CRP protein. Important for targeting TES and ENA/VASP family members to focal adhesions and for the formation of actin-rich structures. May be a component of a signal transduction pathway that mediates adhesion-stimulated changes in gene expression. -
配列類似性
Belongs to the zyxin/ajuba family.
Contains 3 LIM zinc-binding domains. -
細胞内局在
Cytoplasm. Cytoplasm, cytoskeleton. Nucleus. Cell junction, focal adhesion. Associates with the actin cytoskeleton near the adhesion plaques. Enters the nucleus in the presence of HESX1. - Information by UniProt
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別名
- ESP 2
- ESP2
- HED 2
see all
関連製品
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KO cell lines
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Related Products
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab257809の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 61 kDa.
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特記事項 |
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WB
Use at an assay dependent concentration. Predicted molecular weight: 61 kDa. |
画像
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Lane 1: Wild-type HEK-293T cell lysate, 20 ug
Lane 2: ZYX knockout HEK-293T cell lysate, 20 ug
Lane 3: HeLa cell lysate, 20 ug
Lane 4: Daudi cell lysate, 20 ug
False colour image of Western blot: Anti-Zyxin antibody [ZOL301] staining at 1/1000 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab50391 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 CRISPR-Cas9 edited cell line ab266503 (CRISPR-Cas9 edited cell lysate ab257809). The band observed in the CRISPR-Cas9 edited 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 CRISPR-Cas9 edited 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 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-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.
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Lane 1: Wild-type HEK-293T cell lysate, 20 ug
Lane 2: ZYX knockout HEK-293T cell lysate, 20 ug
Lane 3: HeLa cell lysate, 20 ug
Lane 4: Daudi cell lysate, 20 ug
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.
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Lane 1: Wild-type HEK-293T cell lysate, 20 ug
Lane 2: ZYX knockout HEK-293T cell lysate, 20 ug
Lane 3: HeLa cell lysate, 20 ug
Lane 4: Daudi cell lysate, 20 ug
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.
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Allele-1: 19 bp deletion in exon2
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Allele-2: 1 bp deletion in exon2
プロトコール
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (0)
ab257809 は論文での使用が確認できていません。