Human TSC2 (Tuberin) knockout A549 cell line
- Advanced Validation
- What is this?
Be the first to review this product! Submit a review
|
(0 Publication)
TSC2 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View Alternative Names
FLJ43106, LAM, OTTHUMP00000158940, OTTHUMP00000198394, OTTHUMP00000198395, PPP1R160, Protein phosphatase 1, regulatory subunit 160, TSC complex subunit 2, TSC2_HUMAN, TSC4, TSC4 gene, formerly, TSC4, formerly, Tuberin, Tuberous sclerosis 2, Tuberous sclerosis 2 protein, Tuberous sclerosis 2 protein homolog
- WB
Lab
Western blot - Human TSC2 (Tuberin) knockout A549 cell line (AB277867)
Western blot : Anti-TSC2 antibody [EPR22886-235] (ab255612) 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, ab255612 was shown to bind specifically to TSC2. A band was observed at 200 kDa in wild-type A549 cell lysates with no signal observed at this size in TSC2 knockout cell line. To generate this image, wild-type and TSC2 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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-Tuberin antibody [EPR22886-235] (<a href='/en-us/products/primary-antibodies/tuberin-antibody-epr22886-235-ab255612'>ab255612</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
TSC2 knockout A549 cell lysate at 20 µg
Lane 3:
Wild-type HAP1 cell lysate at 20 µg
Lane 4:
TSC2 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 TSC2 (Tuberin) knockout A549 cell line (AB277867)
Western blot : Anti-TSC2 antibody [EP1107Y] (ab52936) staining at 1/50000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab52936 was shown to bind specifically to TSC2. A band was observed at 200 kDa in wild-type A549 cell lysates with no signal observed at this size in TSC2 knockout cell line. To generate this image, wild-type and TSC2 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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-Tuberin antibody [EP1107Y] (<a href='/en-us/products/primary-antibodies/tuberin-antibody-ep1107y-ab52936'>ab52936</a>) at 1/50000 dilution
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
TSC2 knockout A549 cell lysate at 20 µg
Lane 3:
Wild-type HAP1 cell lysate at 20 µg
Lane 4:
TSC2 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 TSC2 (Tuberin) knockout A549 cell line (AB277867)
56 bp deletion after Gln62 of the WT protein.
Reactivity data
Product details
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
Tuberin helps control cell size and proliferation. It does this by modulating the mTOR signaling pathway. This modulation is essential for maintaining cellular homeostasis and energy balance particularly under conditions of nutrient scarcity. Tuberin as part of the TSC1-TSC2 complex also influences cell cycle progression and protein synthesis. Its functions are vital in preventing abnormal cell growth and division which is particularly important in organs where rapid cell turnover occurs.
Pathways
Tuberin plays a central role in the mTOR signaling pathway which affects cell growth autophagy and metabolism. It directly interacts with hamartin (TSC1) to form the TSC1-TSC2 complex which regulates Rheb activity. By inhibiting Rheb Tuberin helps control the mTORC1 pathway linking it to signals such as growth factors stress energy status and amino acid availability. This coordination is important for the proper balance between cell growth and catabolism.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Target data
Complementary Products
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