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AB255421

人PTK2 (FAK) knockout HEK-293T cell line

Human PTK2 (FAK) knockout HEK-293T cell line

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PTK2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 16 bp deletion in exon 4 and 17 bp deletion in exon 4. 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

p125FAK, Focal adhesion kinase 1, PPP1R71, FAK1, Focal adhesion kinase-related nonkinase, FRNK, Protein phosphatase 1 regulatory subunit 71, FAK, Protein-tyrosine kinase 2, PTK2, FADK 1, pp125FAK

4 Images
Western blot - Human PTK2 (FAK) knockout HEK-293T cell line (AB255421)
  • WB

Lab

Western blot - Human PTK2 (FAK) knockout HEK-293T cell line (AB255421)

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.

All lanes:

Western blot - Anti-FAK antibody [EP695Y] (<a href='/products/primary-antibodies/fak-antibody-ep695y-ab40794'>ab40794</a>) at 1/1000 dilution

Lane 1:

HeLa cell lysate at 20 µg

Lane 2:

A431 cell lysate at 20 µg

Lane 3:

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

Lane 4:

Western blot - Human PTK2 (FAK) knockout HEK-293T cell lysate (<a href='/products/cell-lysates/human-ptk2-fak-knockout-hek-293t-cell-lysate-ab263766'>ab263766</a>) at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Predicted band size: 119 kDa

Observed band size: 119 kDa,37 kDa

false

Western blot - Human PTK2 (FAK) knockout HEK-293T cell line (AB255421)
  • WB

Unknown

Western blot - Human PTK2 (FAK) knockout HEK-293T cell line (AB255421)

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.

All lanes:

Western blot - Anti-FAK antibody [EP1831Y] (<a href='/products/primary-antibodies/fak-antibody-ep1831y-ab76496'>ab76496</a>) at 1/500 dilution

Lane 1:

HeLa cell lysate at 20 µg

Lane 2:

A431 cell lysate at 20 µg

Lane 2:

Western blot - Human PTK2 (FAK) knockout HEK-293T cell line (ab255421)

Lane 3:

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

Lane 4:

PTK2 knockout HEK-293T cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Predicted band size: 119 kDa

Observed band size: 119 kDa,37 kDa

false

Sanger Sequencing - Human PTK2 (FAK) knockout HEK-293T cell line (AB255421)
  • Sanger seq

Unknown

Sanger Sequencing - Human PTK2 (FAK) knockout HEK-293T cell line (AB255421)

Allele-2 : 16 bp deletion in exon 4.

Sanger Sequencing - Human PTK2 (FAK) knockout HEK-293T cell line (AB255421)
  • Sanger seq

Unknown

Sanger Sequencing - Human PTK2 (FAK) knockout HEK-293T cell line (AB255421)

Allele-1 : 17 bp deletion in exon 4

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, 16 bp deletion in exon 4 and 17 bp deletion in exon 4

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搭配使用产品

Primary Antibodies

AB50967

Anti-heavy chain cardiac Myosin antibody [BA-G5]

Western blot - Anti-heavy chain cardiac Myosin antibody [BA-G5] (AB50967)

  • 4
  • 12
Primary Antibodies

AB7046

Anti-Collagen V antibody

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Collagen V antibody (AB7046)

  • 5
  • 11
Primary Antibodies

AB211926

Anti-TIMP1 antibody [EPR18352]

20ul selling size|RabMAb|Recombinant|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIMP1 antibody [EPR18352] (AB211926)

  • 5
  • 10
Primary Antibodies

AB109428

Anti-Fibulin 5 antibody [EPR4506]

RabMAb|Recombinant

Western blot - Anti-Fibulin 5 antibody [EPR4506] (AB109428)

  • 5
  • 1
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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.

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
PTK2
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
推荐的短期储存条件
-196°C
推荐的长期储存条件
-196°C
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补充信息

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

Focal Adhesion Kinase (FAK) also known as Protein Tyrosine Kinase 2 (PTK2) is a non-receptor tyrosine kinase. This protein has a molecular weight of approximately 125 kDa. FAK is expressed at high levels in brain muscle and liver tissues. Mechanically FAK plays a role in cellular adhesion and migration by regulating integrin signaling and cell-extracellular matrix interactions. FAK auto-phosphorylates at tyrosine residue 397 creating a binding site for Src family kinases and promoting downstream signaling pathways.
Biological function summary

Focal Adhesion Kinase participates in the formation of focal adhesions which are complexes that connect the cytoskeleton to the extracellular matrix. The FAK protein functions as an important signaling node in these structures allowing for the assembly of multiprotein signal transduction complexes. FAK also controls cellular processes such as spreading motility and survival. The interaction with proteins such as Src kinases paxillin and talin facilitates its biological roles in cell signaling.

Pathways

Focal Adhesion Kinase engages in the regulation of the MAPK/ERK signaling pathway and the PI3K/AKT pathway. These pathways are instrumental for cell proliferation survival and migration. In these pathways FAK interacts with proteins such as PI3K Grb2 and Sos linking integrin-mediated signals with downstream effects that influence cell behavior and survival.

Altered FAK signaling has ties to cancer progression and metastasis as well as cardiovascular diseases. In cancer the overexpression of FAK and its interaction with proteins like Src and VEGFR can drive tumor growth and angiogenesis. In cardiovascular diseases improper FAK activation can lead to aberrant heart tissue remodeling and associated pathologies. Abnormalities in FAK signaling pathways can therefore contribute significantly to the development and progression of these diseases.
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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

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. <p>1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.<br>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.<br>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.<br>4. Incubate the culture at 37°C incubator with 5% CO<sub>2</sub>. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.<br>5. Once confluent passage into an appropriate flask at a density of 2x10<sup>4</sup> cells/cm<sup>2</sup>. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.</p>

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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产品实验方案

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靶点信息

See full target information PTK2
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