Human ATG14 (ATG14L) knockout HeLa cell lysate
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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.
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 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] (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 (<a href='/en-us/products/cell-lines/human-atg14-atg14l-knockout-hela-cell-line-ab264684'>ab264684</a>) 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 lysate (AB258319)
Allele-2 : Insertion of the selection cassette in exon 1
- Sanger seq
Unknown
Sanger Sequencing - Human ATG14 (ATG14L) knockout HeLa cell lysate (AB258319)
Allele-1 : 17 bp deletion in exon 1
Reactivity data
Product details
Knockout cell lysate achieved by CRISPR/Cas9.
REACH authorisation
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.
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
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
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Primary Antibodies
AB313621
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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