ATG14 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 17 bp deletion in exon 1 and Insertion of the selection cassette in exon 1.
Images
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 17 bp deletion in exon 1 and Insertion of the selection cassette in exon 1.
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
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ATG14 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 17 bp deletion in exon 1 and Insertion of the selection cassette in exon 1.
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 17 bp deletion in exon 1 and Insertion of the selection cassette in exon 1.
- Disease
- Adenocarcinoma
- Concentration
Properties
- Gene name
- ATG14
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing, Western blot
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 2 US: 2
- Adherent/suspension
- Adherent
- Gender
- Female
Storage
- Shipped at conditions
- Ambient - Can Ship with Ice
- Appropriate short-term storage conditions
- -20°C
- Appropriate long-term storage conditions
- -20°C
Notes
Knockout cell lysate achieved by CRISPR/Cas9.
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
ATG14L also known as BARKOR is an essential protein involved in the formation of autophagosomes organelles which sequester cytoplasmic components for degradation. This protein is approximately 55 kDa in mass. ATG14L is expressed in a variety of tissues including high expression in the brain heart and kidney. It plays an important role in mediating the recruitment of the PI3K (phosphatidylinositol 3-kinase) complex to the isolation membrane a step necessary in autophagy initiation.
- 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.
- Associated diseases and disorders
ATG14L has a significant connection to neurodegenerative diseases and cancer. Altered autophagy with ATG14L malfunction contributes to the progression of neurodegenerative diseases like Alzheimer's disease where abnormal protein aggregates are not efficiently degraded. In the context of cancer dysregulation of ATG14L-related autophagy pathways can either promote or inhibit tumor growth making it an important therapeutic target. It shares complex interactions with Beclin 1 in these disorders further emphasizing its importance in disease mechanisms.
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3 product images
Western blot - Human ATG14 (ATG14L) knockout HeLa cell lysate (ab258319)
Lane 1: Wild-type HeLa cell lysate 20 μg
Lane 2: HeLa cell lysate 20 μg
Lane 3: HCT 116 cell lysate 20 μg
Lane 4: HepG2 cell lysate 20 μg
False colour image of Western blot: Anti-ATG14 antibody staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control 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 Human ATG14 (ATG14L) knockout HeLa 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed 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 (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
Performed under reducing conditions.
Sanger Sequencing - Human ATG14 (ATG14L) knockout HeLa cell lysate (ab258319)
Allele-2: Insertion of the selection cassette in exon 1
Sanger Sequencing - Human ATG14 (ATG14L) knockout HeLa cell lysate (ab258319)
Allele-1: 17 bp deletion in exon 1
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