Human LAMTOR1 knockout HeLa cell line
- Advanced Validation
- What is this?
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- WB
Lab
Western blot - Human LAMTOR1 knockout HeLa cell line (AB265351)
Lanes 1 - 4 : Merged signal (red and green). Green - ab121157 observed at 18 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
ab121157 was shown to react with LAMTOR1 in wild-type HeLa cells in Western blot with loss of signal observed in LAMTOR1 knockout cell line ab265351 (LAMTOR1 knockout cell lysate ab258938). Wild-type HeLa and LAMTOR1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab121157 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at 0.04 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
Lanes 1 - 4:
Western blot - Anti-LAMTOR1 antibody (<a href='/en-us/products/primary-antibodies/lamtor1-antibody-ab121157'>ab121157</a>) at 0.04 µg/mL
Lanes 1 - 4:
Western blot - Anti-LAMTOR1 antibody (<a href='/en-us/products/primary-antibodies/lamtor1-antibody-ab229760'>ab229760</a>) at 1/500 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
LAMTOR1 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human LAMTOR1 knockout HeLa cell line (ab265351)
Lane 3:
PANC-1 cell lysate at 20 µg
Lane 4:
U-2 OS cell lysate at 20 µg
Predicted band size: 18 kDa
Observed band size: 18 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human LAMTOR1 knockout HeLa cell line (AB265351)
Allele-2 : 4 bp insertion in exon 1.
- Sanger seq
Unknown
Sanger Sequencing - Human LAMTOR1 knockout HeLa cell line (AB265351)
Allele-1 : 1 bp deletion in exon 1.
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Primary Antibodies
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RabMAb|Recombinant
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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, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and 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
- 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.
Culture medium
DMEM (High Glucose) + 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
LAMTOR1 acts as a part of the highly-conserved Ragulator complex. The Ragulator complex is vital for the activation of the mechanistic target of rapamycin complex 1 (mTORC1) in response to amino acids. LAMTOR1's interaction with other components of the complex such as LAMTOR2 and LAMTOR3 facilitates the localization and activation of mTORC1 on the lysosomal surface. This activation promotes anabolic processes including protein lipid and nucleotide synthesis.
Pathways
LAMTOR1 plays an important role in the mTOR signaling pathway. This pathway regulates cell growth proliferation and metabolism. LAMTOR1's involvement with Rag proteins assists in sensing amino acid levels thereby influencing the mTORC1 pathway activation. Additionally LAMTOR1-related pathways intersect with the AMPK signaling pathway where it functionally interacts with proteins that control cellular energy homeostasis.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
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'.
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