Human FTO knockout MCF7 cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Supplier Data
Western blot - Human FTO knockout MCF7 cell line (AB282631)
All lanes:
Western blot - Anti-FTO antibody [EPR6895] (<a href='/en-us/products/primary-antibodies/fto-antibody-epr6895-ab124892'>ab124892</a>) at 1/1000 dilution
Lane 1:
Wild-type MCF7 cell lysate at 20 µg
Lane 2:
FTO knockout MCF7 cell lysate at 20 µg
Lane 2:
Western blot - Human FTO knockout MCF7 cell line (ab282631)
Lane 3:
HEK-293 cell lysate at 20 µg
Lane 4:
MOLT-4 cell lysate at 20 µg
false
- WB
Lab
Western blot - Human FTO knockout MCF7 cell line (AB282631)
All lanes:
Western blot - Anti-FTO antibody [EPR6895] (<a href='/en-us/products/primary-antibodies/fto-antibody-epr6895-ab124892'>ab124892</a>) at 1/1000 dilution
Lane 1:
Wild-type MCF7 cell lysate at 20 µg
Lane 2:
FTO knockout MCF7 cell lysate at 20 µg
Lane 2:
Western blot - Human FTO knockout MCF7 cell line (ab282631)
Lane 3:
HEK-293 cell lysate at 20 µg
Lane 4:
MOLT-4 cell lysate at 20 µg
Predicted band size: 58 kDa
Observed band size: 58 kDa
false
- WB
Supplier Data
Western blot - Human FTO knockout MCF7 cell line (AB282631)
All lanes:
Western blot - Anti-FTO antibody [EPR6894] (<a href='/en-us/products/primary-antibodies/fto-antibody-epr6894-ab126605'>ab126605</a>) at 1/1000 dilution
Lane 1:
Wild-type MCF7 cell lysate at 20 µg
Lane 2:
Western blot - Human FTO knockout MCF7 cell line (ab282631)
Lane 2:
FTO knockout MCF7 cell lysate at 20 µg
Lane 3:
HEK-293 cell lysate at 20 µg
Lane 4:
MOLT-4 cell lysate at 20 µg
Secondary
All lanes:
Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Predicted band size: 58 kDa
Observed band size: 58 kDa
false
- NGS
Lab
Next Generation Sequencing - Human FTO knockout MCF7 cell line (AB282631)
136 bp deletion after the 81st AA of the WT protein
Complementary Products
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FTO antibody [EPR6894] (AB126605)](https://content.abcam.com/products/images/fto-antibody-epr6894-ab126605--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img88351.jpg)
Reactivity data
Product details
We will provide viable cells that proliferate on revival.
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.
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
- Slow to trypsinise.
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 5-7x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
MEM + 10% FBS + 0.01 mg/ml bovine insulin
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
FTO influences energy homeostasis and adipogenesis. It acts as an important player in metabolic regulation and gene expression by modulating the m6A RNA modification. While FTO operates largely on its own its demethylation activity may interact with other RNA binding proteins that recognize m6A-modified RNAs integrating it into larger regulatory frameworks for RNA metabolism and cellular responses.
Pathways
FTO is an important component in the regulation of energy balance and metabolism. The protein is closely associated with the mTOR signaling pathway which is essential for controlling cell growth proliferation and survival in response to nutrient availability. FTO also interacts with the leptin signaling pathway relating to its role in appetite regulation and energy expenditure.
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Female
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