Human PTK2B knockout HCT116 cell line
- KO Validated
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Lab
Western blot - Human PTK2B knockout HCT116 cell line (AB301190)
Western blot : Rabbit Monoclonal[EP206Y] to PYK2 ab81266 staining at 1/2500 dilution, shown in green; Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta.
A band was observed at 120 kDa in Wild-type HCT 116 cell lysates with no signal observed at this size in PTK2B knockout HCT 116 cell line (ab301190). We are unable to identify the non-specific bands at 75kDa and 40kDa.
To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc 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 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.
All lanes:
Western blot - Anti-PYK2 antibody [EP206Y] (<a href='/en-us/products/primary-antibodies/pyk2-antibody-ep206y-ab81266'>ab81266</a>) at 1/1000 dilution
Lane 1:
Wild-type HCT 116 at 20 µg
Lane 2:
Western blot - Human PTK2B knockout HCT116 cell line (ab301190) at 20 µg
Lane 3:
Wild-type A549 at 20 µg
Lane 4:
PTK2B knockout A549 at 20 µg
Secondary
All lanes:
Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution
Predicted band size: 116 kDa
Observed band size: 120 kDa
false
- WB
Lab
Western blot - Human PTK2B knockout HCT116 cell line (AB301190)
Western blot : Rabbit Monoclonal[YE353] to PYK2 ab32571 staining at 1/2000 dilution, shown in green; Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta.
A band was observed at 120 kDa in Wild-type HCT 116 cell lysates with no signal observed at this size in PTK2B knockout HCT 116 cell line (ab301190).
To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc 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 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.
All lanes:
Western blot - Anti-PYK2 antibody [YE353] (<a href='/en-us/products/primary-antibodies/pyk2-antibody-ye353-ab32571'>ab32571</a>) at 1/1000 dilution
Lane 1:
Wild-type HCT 116 at 20 µg
Lane 2:
Western blot - Human PTK2B knockout HCT116 cell line (ab301190) at 20 µg
Lane 3:
Wild-type A549 at 20 µg
Lane 4:
PTK2B knockout A549 at 20 µg
Secondary
All lanes:
Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution
Predicted band size: 116 kDa
Observed band size: 120 kDa
false
- NGS
Lab
Next Generation Sequencing - Human PTK2B knockout HCT116 cell line (AB301190)
106 bp deletion after Val33 (edit 1); 73 bp deletion after Lys16 (edit 2)
- WB
Lab
Western blot - Human PTK2B knockout HCT116 cell line (AB301190)
Western blot : Rabbit Monoclonal [EP206Y] to PYK2 ab81266 staining at 1/2000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 116 kDa in Wild-type HCT 116 cell lysates with no signal observed at this size in PTK2B knockout HCT 116 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5pc 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 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.
All lanes:
Western blot - Anti-PYK2 antibody [EP206Y] (<a href='/en-us/products/primary-antibodies/pyk2-antibody-ep206y-ab81266'>ab81266</a>) at 1/2000 dilution
Lane 1:
Wild-type HCT 116 at 20 µg
Lane 2:
Western blot - Human PTK2B knockout HCT116 cell line (ab301190) at 20 µg
Lane 3:
A549 at 20 µg
Secondary
All lanes:
Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution
Predicted band size: 116 kDa
Observed band size: 116 kDa
false
Complementary Products
Primary Antibodies
AB32571
RabMAb|Recombinant|KO Validated|20ul selling size
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PYK2 antibody [YE353] (AB32571)](https://content.abcam.com/products/images/pyk2-antibody-ye353-ab32571--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img129863.jpg)
- 0
- 0
Reactivity data
Product details
Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.
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
McCoY5a + 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
PYK2 interacts with integrins and G-protein-coupled receptors (GPCRs) linking extracellular signals to intracellular pathways. It forms complexes with other signaling molecules including Rac1 and c-Src facilitating downstream signaling. These interactions contribute to cellular communication and adhesion processes especially in the central nervous system and immune cells. Through its involvement in signal transduction PYK2 supports cellular responses to environmental changes.
Pathways
PYK2 participates in the MAPK and JNK signaling pathways. These pathways are critical for transmitting signals from the cell surface to the nucleus influencing cellular growth and stress responses. PYK2 interacts closely with other proteins such as Src and FAK which help coordinate signals that regulate cellular dynamics. Through these pathways PYK2 modulates processes involved in the structural reorganization of cells and tissues.
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Target data
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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