Human TNK2 knockout HCT116 cell line
- KO Validated
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Lab
Western blot - Human TNK2 knockout HCT116 cell line (AB301274)
Western blot : anti-ACK antibody staining at 1/1000 dilution, shown in green; Rabbit anti alpha Tubulin (ab52866) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 118 kDa in Wild-type HCT 116 cell lysates with no signal observed at this size in TNK2 knockout HCT 116 cell line (ab301274). 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-Mouse 800CW & Goat anti-Rabbit 680RD at 1/20,000 dilution.
All lanes:
Western blot at 1/1000 dilution
Lane 1:
Wild-type HCT 116 at 20 µg
Lane 2:
Western blot - Human TNK2 knockout HCT116 cell line (ab301274) at 20 µg
Lane 3:
HeLa at 20 µg
Lane 4:
PC-3
Secondary
All lanes:
Goat anti-Mouse 800CW & Goat anti-Rabbit 680RD at 1/20000 dilution
false
- NGS
Lab
Next Generation Sequencing - Human TNK2 knockout HCT116 cell line (AB301274)
7 bp deletion and 115 bp deletion after Ala271
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
The functions of ACK1 relate to signal transduction processes. It acts as an important part of the signaling complex involved in cell growth and differentiation. ACK1 interacts with several growth factor receptors and coordinates signal transduction events. These interactions allow ACK1 to participate in complex networks that regulate cellular shaping and organization making it important for normal cell functionality.
Pathways
ACK1 significantly influences cellular processes through the MAPK and PI3K pathways. These pathways are essential for cell survival proliferation and metabolism. ACK1 interacts with proteins such as ERK and AKT mediating downstream signaling that determines cellular responses to external stimuli. The kinase activity of ACK1 regulates various signaling cascades emphasizing its role in controlling metabolic and growth signals.
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'.
For licensing inquiries, please contact partnerships@abcam.com