Human RICTOR knockout A549 cell line
- Advanced Validation
- What is this?
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RICTOR KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 43 bp deletion in exon 5.
View Alternative Names
AVO3, AVO3 homolog, DKFZp686B11164, KIAA1999, Likely ortholog of mouse TORC2 specific protein AVO3 (S. cerevisiae), MGC39830, PIA, Pianissimo, RICTR_HUMAN, RPTOR independent companion of MTOR complex 2, Rapamycin-insensitive companion of mTOR, TORC2 specific protein AVO3, hAVO3, mAVO3
- WB
Lab
Western blot - Human RICTOR knockout A549 cell line (AB277866)
False colour image of Western blot : Anti-RICTOR antibody [EPR22008] staining at 1/1000 dilution shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution shown in red. In Western blot ab219950 was shown to bind specifically to RICTOR. A band was observed at 190 kDa in wild-type A549 cell lysates with no signal observed at this size in RICTOR knockout cell line ab277866 (knockout cell lysate ab288315). To generate this image wild-type and RICTOR 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 °. 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-RICTOR antibody [EPR22008] (<a href='/en-us/products/primary-antibodies/rictor-antibody-epr22008-ab219950'>ab219950</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
RICTOR knockout A549 cell lysate at 20 µg
Lane 2:
Western blot - Human RICTOR knockout A549 cell line (ab277866)
Lane 3:
HeLa cell lysate at 20 µg
Predicted band size: 192 kDa
Observed band size: 190 kDa
false
- WB
Lab
Western blot - Human RICTOR knockout A549 cell line (AB277866)
False colour image of Western blot : Anti-RICTOR antibody staining at 1 ug/ml shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution shown in red. In Western blot ab105469 was shown to bind specifically to RICTOR. A band was observed at 190 kDa in wild-type A549 cell lysates with no signal observed at this size in RICTOR knockout cell line ab277866 (knockout cell lysate ab288315). To generate this image wild-type and RICTOR 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 °. 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-RICTOR antibody (<a href='/en-us/products/primary-antibodies/rictor-antibody-ab105469'>ab105469</a>) at 1 µg/mL
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
RICTOR knockout A549 cell lysate at 20 µg
Lane 2:
Western blot - Human RICTOR knockout A549 cell line (ab277866)
Lane 3:
HeLa cell lysate at 20 µg
Predicted band size: 192 kDa
Observed band size: 190 kDa
false
- Sanger seq
Supplier Data
Sanger Sequencing - Human RICTOR knockout A549 cell line (AB277866)
43 bp deletion in exon 5
Reactivity data
Product details
Recommended control: Human wild-type A549 cell line (ab255450). 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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Zygosity
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
The protein plays a significant role in cellular growth proliferation and survival. RICTOR forms part of the mTORC2 complex which is important for Akt/PKB phosphorylation. Activation of mTORC2 by RICTOR regulates actin cytoskeleton dynamics and membrane trafficking. This involvement affects the control of cell metabolism and immune responses.
Pathways
RICTOR integrates into the PI3K/Akt signaling pathway influencing cell growth and survival in response to nutrients and growth factors. RICTOR as part of mTORC2 also interacts with proteins such as Sin1 and mLST8 helping regulate PKB/Akt phosphorylation at Ser473. This interaction links RICTOR's function to the insulin signaling pathway which controls aspects of glucose metabolism.
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'.
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