Human LAMTOR2 knockout HeLa cell line
- Advanced Validation
- What is this?
Be the first to review this product! Submit a review
|
(0 Publication)
LAMTOR2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 2 and 7 bp deletion in exon 2.
View Alternative Names
ENDAP, Endosomal adaptor protein p14, HSPC003, LTOR2_HUMAN, Late endosomal/lysosomal Mp1-interacting protein, Late endosomal/lysosomal adaptor and MAPK and MTOR activator 2, MAPBPIP, MAPKSP1 adaptor protein, MAPKSP1AP, Mitogen-activated protein-binding protein-interacting protein, P14, ROBLD 3, RP11 336K24.9, Ragulator complex protein LAMTOR2, Ragulator2, Roadblock domain containing 3, Roadblock domain-containing protein 3
- WB
Lab
Western blot - Human LAMTOR2 knockout HeLa cell line (AB265828)
Lanes 1-4 : Merged signal (red and green). Green - ab183514 observed at 14 kDa. Red - loading control ab8245 observed at 37 kDa.
ab183514 Anti-LAMTOR2 antibody [EPR14378] was shown to specifically react with LAMTOR2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265828 (knockout cell lysate ab258020) was used. Wild-type and LAMTOR2 knockout samples were subjected to SDS-PAGE. ab183514 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-LAMTOR2 antibody [EPR14378] (<a href='/en-us/products/primary-antibodies/lamtor2-antibody-epr14378-ab183514'>ab183514</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
LAMTOR2 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human LAMTOR2 knockout HeLa cell line (ab265828)
Lane 3:
HepG2 cell lysate at 20 µg
Lane 4:
LNCaP cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution
Predicted band size: 14 kDa
Observed band size: 14 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human LAMTOR2 knockout HeLa cell line (AB265828)
Allele-2 : 1 bp deletion in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human LAMTOR2 knockout HeLa cell line (AB265828)
Allele-1 : 7 bp deletion in exon 2.
Reactivity data
Product details
Recommended control: Human wild-type HeLa cell line (ab255928). 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
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
LAMTOR2 serves in the regulation of mTORC1 signaling which plays a major role in cell growth and metabolism. As a member of the Ragulator complex LAMTOR2 works in concert with other proteins like LAMTOR1 and LAMTOR3 to anchor the Rag GTPases to the lysosomal surface. This provides a platform for the activation of mTORC1 in response to amino acid availability therefore guiding cellular energy and protein synthesis pathways.
Pathways
LAMTOR2 functions integrally in the amino acid-sensing pathway and impacts the mTOR signaling pathway. The Ragulator complex involving LAMTOR2 interacts with Rag GTPases to activate mTORC1 upon amino acid availability bridging nutrient sensing and cellular metabolism control. In doing so it interacts closely with proteins like RagA/RagB coordinating signals that are critical for growth-related cellular functions.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Complementary Products
![Western blot - Anti-LAMTOR2 antibody [EPR14378] (AB183514)](https://content.abcam.com/products/images/lamtor2-antibody-epr14378-ab183514--western-blot-img71369.jpg)
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