FTO KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided.
Images
Key facts
- Cell type
- MCF7
- Species or organism
- Human
- Tissue
- Breast
- Form
- Liquid
- Knockout validation
- Western blot
Alternative names
ALKBH9, AW743446, AlkB homolog 9, Alpha-ketoglutarate-dependent dioxygenase FTO, FATSO, MOUSE, HOMOLOG OF, FTO_HUMAN, Fat mass and obesity-associated protein, GDFD, KIAA1752, Protein fatso, mKIAA1752
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FTO KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided.
Key facts
- Cell type
- MCF7
- Species or organism
- Human
- Tissue
- Breast
- Form
- Liquid
- Knockout validation
- Western blot
- Disease
- Adenocarcinoma
- Concentration
Properties
- Gene name
- FTO
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Western blot
Cell culture
- Biosafety level
- EU: 1 US: 1
- Adherent/suspension
- Adherent
- Gender
- Female
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.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
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 MCF7 cell line (ab288560). 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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
The FTO protein also referred to as Alpha-Ketoglutarate-Dependent Dioxygenase or Fat Mass and Obesity-Associated protein functions as an enzyme involved in demethylating nucleic acids specifically N6-methyladenosine (m6A) in RNA. This process impacts RNA stability and translation. FTO has a molecular weight of approximately 58 kDa. Higher expression levels of FTO are found in the brain including the hypothalamus which is an area known for regulating energy balance and appetite.
- 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.
- Associated diseases and disorders
FTO has significant connections to obesity and Type 2 diabetes. Genetic variants within the FTO gene correlate with risk for these conditions linking its enzymatic activity to alterations in energy balance and insulin sensitivity. FTO's role in Type 2 diabetes also highlights its interaction with insulin signaling proteins demonstrating its broader influence on glucose metabolism.
Product promise
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
4 product images
Western blot - Human FTO knockout MCF7 cell line (ab282631)
All lanes: Western blot - Anti-FTO antibody [EPR6895] (Anti-FTO antibody [EPR6895] ab124892) 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
Performed under reducing conditions.
Predicted band size: 58 kDa
Observed band size: 58 kDa
Western blot - Human FTO knockout MCF7 cell line (ab282631)
All lanes: Western blot - Anti-FTO antibody [EPR6895] (Anti-FTO antibody [EPR6895] ab124892) 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
Next Generation Sequencing - Human FTO knockout MCF7 cell line (ab282631)
136 bp deletion after the 81st AA of the WT protein
Western blot - Human FTO knockout MCF7 cell line (ab282631)
All lanes: Western blot - Anti-FTO antibody [EPR6894] (Anti-FTO antibody [EPR6894] ab126605) 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
SecondaryAll lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 58 kDa
Observed band size: 58 kDa
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