人PTK2 (FAK) knockout HEK-293T cell line (ab255421)
概述
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产品名称
人PTK2 (FAK) knockout HEK-293T cell line
参阅全部 FAK 细胞裂解液 -
Parental Cell Line
HEK293T -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, 16 bp deletion in exon 4 and 17 bp deletion in exon 4 -
Passage number
<20 -
Knockout validation
Sanger Sequencing, Western Blot (WB) -
经测试应用
适用于: WBmore details -
Biosafety level
2 -
常规说明
Recommended control: Human wild-type HEK293T cell line (ab255449). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
Cryopreservation cell medium: Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Culture medium: DMEM (High Glucose) + 10% FBS
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. Based on cell count, seed cells in an appropriate cell culture 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.
4. Incubate the culture at 37°C incubator with 5% CO2. 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.
- A partial media change 24 hours prior to subculture may be helpful to encourage growth, if required.
- Cells should be passaged when they have achieved 80-90% confluence.
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.
We will provide viable cells that proliferate on revival.
性能
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Number of cells
1 x 106 cells/vial, 1 mL -
Adherent /Suspension
Adherent -
Tissue
Kidney -
Cell type
epithelial -
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 -
Mycoplasma free
Yes -
存放说明
Shipped on Dry Ice. Store in liquid nitrogen. -
存储溶液
Constituents: 8.7% Dimethylsulfoxide, 2% Cellulose, methyl ether -
研究领域
靶标
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功能
Non-receptor protein-tyrosine kinase implicated in signaling pathways involved in cell motility, proliferation and apoptosis. Activated by tyrosine-phosphorylation in response to either integrin clustering induced by cell adhesion or antibody cross-linking, or via G-protein coupled receptor (GPCR) occupancy by ligands such as bombesin or lysophosphatidic acid, or via LDL receptor occupancy. Microtubule-induced dephosphorylation at Tyr-397 is crucial for the induction of focal adhesion disassembly. Plays a potential role in oncogenic transformations resulting in increased kinase activity. -
组织特异性
Expressed in all organs tested, in lymphoid cell lines, but most abundantly in brain. -
序列相似性
Belongs to the protein kinase superfamily. Tyr protein kinase family. FAK subfamily.
Contains 1 FERM domain.
Contains 1 protein kinase domain. -
结构域
The first Pro-rich domain interacts with the SH3 domain of CRK-associated substrate (BCAR1) and CASL.
The carboxy-terminal region is the site of focal adhesion targeting (FAT) sequence which mediates the localization of FAK1 to focal adhesions. -
翻译后修饰
Phosphorylated on 6 tyrosine residues upon activation. Microtubule-induced dephosphorylation at Tyr-397 could be catalyzed by PTPN11 and regulated by ZFYVE21. Dephosphorylated by PTPN11 upon EPHA2 activation by its ligand EFNA1. -
细胞定位
Cell junction > focal adhesion. Cell membrane. Constituent of focal adhesions. - Information by UniProt
相关产品
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KO cell lysates
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab255421于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 119 kDa.
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说明 |
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WB
Use at an assay dependent concentration. Predicted molecular weight: 119 kDa. |
图片
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All lanes : Anti-FAK antibody [EP1831Y] (ab76496) at 1/500 dilution
Lane 1 : HeLa cell lysate
Lane 2 : A431 cell lysate
Lane 3 : Wild-type HEK-293T cell lysate
Lane 4 : PTK2 knockout HEK-293T cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 119 kDa
Additional bands at: 37 kDa (possible Loading Control)Lanes 1 - 4: Merged signal (red and green). Green - ab76496 observed at 119 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab76496 was shown to react with FAK in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab255421 (knockout cell lysate ab263766) was used. Wild-type and FAK knockout samples were subjected to SDS-PAGE. ab76496 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-FAK antibody [EP695Y] (ab40794) at 1/1000 dilution
Lane 1 : HeLa cell lysate
Lane 2 : A431 cell lysate
Lane 3 : Wild-type HEK-293T cell lysate
Lane 4 : PTK2 knockout HEK-293T cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 119 kDa
Additional bands at: 37 kDa (possible Loading Control)Lanes 1 - 4: Merged signal (red and green). Green - ab40794 observed at 119 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab40794 was shown to react with FAK in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab255421 (knockout cell lysate ab263766) was used. Wild-type and FAK knockout samples were subjected to SDS-PAGE. ab40794 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Allele-1: 17 bp deletion in exon 4
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Allele-2: 16 bp deletion in exon 4.
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (0)
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