Human PTK2 (FAK) knockout HEK-293T cell line
- Advanced Validation
- What is this?
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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'.
View Alternative Names
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
- 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='/en-us/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='/en-us/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
- 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='/en-us/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 (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='/en-us/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 seq
Unknown
Sanger Sequencing - Human PTK2 (FAK) knockout HEK-293T cell line (AB255421)
Allele-2 : 16 bp deletion in exon 4.
- Sanger seq
Unknown
Sanger Sequencing - Human PTK2 (FAK) knockout HEK-293T cell line (AB255421)
Allele-1 : 17 bp deletion in exon 4
Reactivity data
Product details
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Handling procedures
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 seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate 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. 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.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
Primary Antibodies
AB81298
Anti-FAK (phospho Y397) antibody [EP2160Y]
20ul selling size|RabMAb|Recombinant
![Western blot - Anti-FAK (phospho Y397) antibody [EP2160Y] (AB81298)](https://content.abcam.com/products/images/fak-phospho-y397-antibody-ep2160y-ab81298--western-blot-img149070.jpg)
- 4
- 2
Product promise
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