Human ATG14 (ATG14L) knockout HeLa cell line
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ATG14 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 17 bp deletion in exon 1 and Insertion of the selection cassette in exon 1. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View Alternative Names
4832427M01, ATG14, Autophagy-related protein 14-like protein, BAKOR_HUMAN, Barkor, Beclin 1-associated autophagy-related key regulator, D14Ertd114e, D14Ertd436e, KIAA0831, mCG_6911
- WB
Lab
Western blot - Human ATG14 (ATG14L) knockout HeLa cell line (AB264684)
False colour image of Western blot : Anti-ATG14 antibody staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, the antibody was shown to bind specifically to ATG14. A band was observed at 55 kDa in wild-type HeLa cell lysates with no signal observed at this size in ATG14 knockout cell line ab264684 (knockout cell lysate ab258319). To generate this image, wild-type and ATG14 knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Anti-ATG14 antibody at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human ATG14 (ATG14L) knockout HeLa cell line (ab264684) at 20 µg
Lane 3:
HCT 116 cell lysate at 20 µg
Lane 4:
HepG2 cell lysate at 20 µg
false
- Sanger seq
Unknown
Sanger Sequencing - Human ATG14 (ATG14L) knockout HeLa cell line (AB264684)
Allele-2 : Insertion of the selection cassette in exon 1.
- Sanger seq
Unknown
Sanger Sequencing - Human ATG14 (ATG14L) knockout HeLa cell line (AB264684)
Allele-1 : 17 bp deletion in exon 1.
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
ATG14L coordinates with other proteins as part of the PI3K complex performing a central function in the regulation of autophagy the cellular degradation process critical for maintaining cellular homeostasis. This complex includes VPS34 VPS15 and Beclin 1 which work together to facilitate the nucleation of the autophagosome membrane. Autophagy driven by ATG14L is important for cellular responses to nutrient deprivation and stress.
Pathways
ATG14L is important for two central pathways: the regulation of autophagy and the PI3K/AKT/mTOR signaling pathways. ATG14L directly affects the activity of mTORC1 an important regulator of cell growth and metabolism integrating signals from nutrients and growth factors. It interacts with other autophagy-related proteins such as Beclin 1 to modulate these pathways connecting intracellular energy status with metabolic processes.
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
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