Human STK11 (LKB1) knockout HEK-293T cell line
- Advanced Validation
- What is this?
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- WB
Lab
Western blot - Human STK11 (LKB1) knockout HEK-293T cell line (AB266647)
Lanes 1- 4 : Merged signal (red and green). Green - ab199970 observed at 55 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab199970 was shown to react with LKB1 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266647 (knockout cell lysate ab257712) was used. Wild-type HEK-293T and STK11 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab199970 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4° at a 1 in 1000 dilution and a 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-LKB1 antibody [EPR19379] (<a href='/en-us/products/primary-antibodies/lkb1-antibody-epr19379-ab199970'>ab199970</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
STK11 knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human STK11 (LKB1) knockout HEK-293T cell line (ab266647)
Lane 3:
Hu testis cell lysate at 20 µg
Lane 4:
HeLa cell lysate at 20 µg
Predicted band size: 48 kDa
Observed band size: 55 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human STK11 (LKB1) knockout HEK-293T cell line (AB266647)
Homozygous : 1 bp insertion in exon1
Complementary Products
![Immunoprecipitation - Anti-LKB1 antibody [EPR19379] (AB199970)](https://content.abcam.com/products/images/lkb1-antibody-epr19379-ab199970--immunoprecipitation-img64560.jpg)
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
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.
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
LKB1 is involved in controlling cell growth polarity and metabolism. It forms a complex with STRAD and MO25 which are necessary for its activation and function. By influencing these biological processes LKB1 regulates cell cycle arrest in response to stress signals and contributes to maintaining cellular integrity. The regulation of these processes positions LKB1 as an important factor in managing how cells respond to metabolic challenges.
Pathways
Its function integrates into major signaling networks like the AMPK and mTOR pathways. LKB1 activates AMPK which in turn affects mTOR signaling a pathway critical for cell growth and autophagy regulation. LKB1's interaction with AMPK links cellular energy levels to the control of biosynthetic pathways and cell cycle progression. The LKB1-AMPK-mTOR axis for instance highlights the protein's role in managing how cells balance growth-related activities and energy conservation.
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'.
For licensing inquiries, please contact partnerships@abcam.com