Human TSC1 knockout MCF7 cell line
- Advanced Validation
- What is this?
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- WB
Lab
Western blot - Human TSC1 knockout MCF7 cell line (AB286332)
Western blot : Anti-TSC1 antibody [EP318Y] (ab40872) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab40872 was shown to bind specifically to TSC1. A band was observed at 130-150 kDa in wild-type MCF7 cell lysates with no signal observed at this size in TSC1 knockout cell line. To generate this image, wild-type and TSC1 knockout MCF7 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % 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 IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes:
Western blot - Anti-Hamartin antibody [EP318Y] (<a href='/en-us/products/primary-antibodies/hamartin-antibody-ep318y-ab40872'>ab40872</a>) at 1/1000 dilution
Lane 1:
Wild-type MCF7 cell lysate at 20 µg
Lane 2:
TSC1 knockout MCF7 cell lysate at 20 µg
Lane 3:
Wild-type A549 cell lysate at 20 µg
Lane 4:
TSC1 knockout A549 cell lysate at 20 µg
Lane 5:
Wild-type HAP1 cell lysate at 20 µg
Lane 6:
TSC1 knockout HAP1 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
false
- WB
Lab
Western blot - Human TSC1 knockout MCF7 cell line (AB286332)
Western blot : Anti-TSC1 antibody [EPR24364-109] (ab270967) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab270967 was shown to bind specifically to TSC1. A band was observed at 140-150 kDa in wild-type MCF7 cell lysates with no signal observed at this size in TSC1 knockout cell line. To generate this image, wild-type and TSC1 knockout MCF7 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % 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 IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes:
Western blot - Anti-Hamartin antibody [EPR24364-109] (<a href='/en-us/products/primary-antibodies/hamartin-antibody-epr24364-109-ab270967'>ab270967</a>) at 1/1000 dilution
Lane 1:
Wild-type MCF7 cell lysate at 20 µg
Lane 2:
TSC1 knockout MCF7 cell lysate at 20 µg
Lane 3:
Wild-type A549 cell lysate at 20 µg
Lane 4:
TSC1 knockout A549 cell lysate at 20 µg
Lane 5:
Wild-type HAP1 cell lysate at 20 µg
Lane 6:
TSC1 knockout HAP1 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
false
- NGS
Supplier Data
Next Generation Sequencing - Human TSC1 knockout MCF7 cell line (AB286332)
Various deletions of about 191 bp after Gly527 of the WT protein
Complementary Products
Reactivity data
Product details
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
- 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
TSC-1 plays a regulatory role in cell growth and proliferation. It forms a complex with TSC-2 also known as tuberin which is important for its function. This complex acts as a GAP (GTPase-activating protein) toward Rheb a small GTPase inhibiting the mTOR signaling pathway. The proper regulation of this pathway ensures controlled cell size and growth emphasizing the importance of the TSC-1/TSC-2 complex in cellular regulation.
Pathways
TSC-1 operates within the mTOR signaling pathway to influence cellular processes like autophagy and protein synthesis. It also relates to the PI3K/AKT pathway one that is involved in the regulation of cell survival and metabolism. By interacting with TSC-2 in these pathways TSC-1 modulates the inhibition of mTORC1 thereby impacting cellular homeostasis and growth.
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