AB263766

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

Name: Human PTK2 (FAK) knockout HEK-293T cell lysate (ab263766) | Abcam
Brand: Abcam Limited
SKU: AB263766
Price: 114000 JPY
Availability: InStock

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

別名を表示する

FADK, FADK 1, FAK related non kinase polypeptide, FAK1_HUMAN, FRNK, Focal adhesion Kinase, Focal adhesion kinase 1, Focal adhesion kinase isoform FAK Del33, Focal adhesion kinase related nonkinase, PPP1R71, PTK2 protein tyrosine kinase 2, Protein phosphatase 1 regulatory subunit 71, Protein-tyrosine kinase 2, Ptk2, p125FAK, pp125FAK

3 Images

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Western blot - Human PTK2 (FAK) knockout HEK-293T cell lysate (AB263766)

Lane 1 : HeLa cell lysate (20 μg)

Lane 2 : A431 cell lysate (20 μg)

Lane 3 : Wild-type HEK-293T cell lysate (20 μg)

Lane 4 : PTK2 knockout HEK-293T cell lysate (20 μg)

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 (<a href='/products/cell-lines/human-ptk2-fak-knockout-hek-293t-cell-line-ab255421'>ab255421</a>)

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

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Lab

Western blot - Human PTK2 (FAK) knockout HEK-293T cell lysate (AB263766)

Lane 1 : HeLa cell lysate (20 μg)

Lane 2 : A431 cell lysate (20 μg)

Lane 3 : Wild-type HEK-293T cell lysate (20 μg)

Lane 4 : PTK2 knockout HEK-293T cell lysate (20 μg)

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 2:

Western blot - Human PTK2 (FAK) knockout HEK-293T cell line (<a href='/products/cell-lines/human-ptk2-fak-knockout-hek-293t-cell-line-ab255421'>ab255421</a>)

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 lysate (AB263766)

Sanger seq

Unknown

Sanger Sequencing - Human PTK2 (FAK) knockout HEK-293T cell lysate (AB263766)

Allele-1 : 17 bp deletion in exon 4; Allele-2 : 16 bp deletion in exon 4

Key facts

細胞タイプ

HEK-293T

生物種

Human

組織

Kidney

ノックアウト検証方法

Sanger Sequencing,Western blot

ノックアウト変異

Knockout achieved by using CRISPR/Cas9, 16 bp deletion in exon 4 and 17 bp deletion in exon 4.

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Reactivity data

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製品の詳細

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 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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製品内容

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab263766-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human PTK2 knockout HEK293T cell lysate", "number":"AB263766-CMP01", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type HEK293T cell lysate", "number":"AB263766-CMP02", "productcode":"" } ] } } }

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出荷温度及び保存条件

遺伝子名

PTK2

遺伝子編集のタイプ

Knockout

遺伝子編集の方法

CRISPR technology

ノックアウト検証方法

Sanger Sequencing, Western blot

出荷温度

Ambient - Can Ship with Ice

短期保存温度

-20°C

長期保存温度

-20°C

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補足情報

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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Quality control

STR analysis

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

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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製品プロトコール

Visit the General protocols
Visit the Troubleshooting

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Abcam product promise

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