Human TBX3 knockout A549 cell line
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Human TBX3 knockout A549 cell line available to order. Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein.
View Alternative Names
Bladder cancer related protein XHL, T box 3, T-box protein 3, T-box transcription factor TBX3, TBX3 ISO, TBX3_HUMAN, UMS, XHL
- WB
Lab
Western blot - Human TBX3 knockout A549 cell line (AB301265)
Western blot : TBX3 Polyclonal Antibody ab42-4800 staining at 2 µg/mL, shown in green; Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 79 kDa in Wild-type A549 cell lysates with no signal observed at this size in TBX3 knockout A549 cell line. 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 at 2 µg/mL
Lane 1:
Wild-type A549 at 20 µg
Lane 2:
Western blot - Human TBX3 knockout A549 cell line (ab301265) at 20 µg
Lane 3:
HepG2 at 20 µg
Lane 4:
MOLT-4 at 20 µg
Secondary
All lanes:
Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution
false
- NGS
Lab
Next Generation Sequencing - Human TBX3 knockout A549 cell line (AB301265)
176 bp deletion after Gln 66 (allele 1); 5 bp insertion and 63 bp deletion after Asp 65 (allele 2); 67 bp insertion and 80 bp deletion after Gln 66 (allele 3) of WT protein
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.
Recommended control: Human wild-type A549 cell line (ab288558). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
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.
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.
- Do not allow the cell density to exceed 7x104 cells/cm2.
Culture medium
F-12K + 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
T-box transcription factor 3 is significant in embryonic development and cellular differentiation. It plays an important role in the development of the heart and limbs impacting cell fate decisions. Tbx3 interacts with other T-box family members and it is part of a regulatory network controlling pluripotency and differentiation in stem cells. Its expression patterns suggest important functions in organogenesis and tissue specification.
Pathways
Tbx3 is a modulatory component in pathways controlling development and cell cycle progression. It is closely involved in the Wnt signaling and cell differentiation pathways. In the Wnt pathway Tbx3 regulates genes essential for cell proliferation and development. It interacts with proteins such as β-catenin linking it to other pathway members like LEF/TCF transcription factors further influencing cellular responses and development processes.
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Product protocols
- Visit the General protocols
- Visit the Troubleshooting
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