ATG7 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, functional Homozygous: 41 dp deletion in exon 4.
Images
Key facts
- Cell type
HeLa
- Species or organism
Human
- Tissue
Cervix
- Form
Liquid
- Knockout validation
Sanger Sequencing, Western blot
- Mutation description
Knockout achieved by using CRISPR/Cas9, functional Homozygous: 41 dp deletion in exon 4.
Alternative names
1810013K23Rik, APG7 autophagy 7 like, APG7 autophagy 7-like (S. cerevisiae), APG7, S. cerevisiae, homolog of, APG7-like, APG7L, ATG12-activating enzyme E1 ATG7, ATG7 autophagy related 7 homolog, ATG7 autophagy related 7 homolog (S. cerevisiae), ATG7_HUMAN, Apg 7, Atg7l, Autophagy 7, S. cerevisiae, homolog of, Autophagy-related 7 (yeast), Autophagy-related protein 7, DKFZp434N0735, GSA 7, Ubiquitin-activating enzyme E1-like protein, Ubiquitin-like modifier-activating enzyme ATG7, hAGP7
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ATG7 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, functional Homozygous: 41 dp deletion in exon 4.
Key facts
- Cell type
HeLa
- Species or organism
Human
- Tissue
Cervix
- Form
Liquid
- Knockout validation
Sanger Sequencing, Western blot
- Mutation description
Knockout achieved by using CRISPR/Cas9, functional Homozygous: 41 dp deletion in exon 4.
- Disease
Adenocarcinoma
- Concentration
Properties
- Gene name
ATG7
- Gene editing type
Knockout
- Gene editing method
CRISPR technology
- Knockout validation
Sanger Sequencing, Western blot
- Zygosity
Homozygous
Quality control
- STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
EU: 2 US: 2
- Adherent/suspension
Adherent
- Gender
Female
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.
Storage
- Shipped at conditions
Dry Ice
- Appropriate short-term storage conditions
-196°C
- Appropriate long-term storage conditions
-196°C
Notes
Recommended control: Human wild-type HeLa cell line (Human wild-type HeLa cell line ab275466). 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.
Supplementary info
This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
ATG7 also known as Autophagy Related 7 is an essential protein involved in the autophagy process. It functions as an E1-like enzyme activating and transferring ubiquitin-like proteins such as ATG12 and LC3. The molecular weight of ATG7 is approximately 78 kDa. It is widely expressed in various tissues although expression levels can differ. One can detect ATG7 using immunoassays like ELISA or antibodies specifically targeting ATG7. Understanding its mechanical role is key to studying cellular homeostasis.
- Biological function summary
ATG7 plays a significant role in autophagy a cellular degradation pathway critical for cell survival under stress. ATG7 contributes to the formation of autophagosomes by facilitating conjugation of ATG8 family proteins including LC3 to phosphatidylethanolamine. It acts within complexes that regulate cellular energy balance and stress responses ensuring cells maintain their function and integrity. Knockdown of ATG7 can impair autophagic flux highlighting its importance in maintaining cellular processes.
- Pathways
ATG7 is a central player in the autophagy pathway influencing cellular metabolism and turnover. It interacts closely with ATG5 and ATG12 in this pathway to form a conjugation system essential for autophagosome elongation. Additionally ATG7 is involved in the mTOR signaling pathway which regulates nutrient sensing and cellular growth. Interaction with proteins like mTOR allows ATG7 to integrate signals from nutrient availability and stress responses finely tuning the autophagy process.
- Associated diseases and disorders
ATG7 dysfunction has connections to cancer and neurodegenerative diseases like Alzheimer's. Abnormal ATG7 activity disrupts autophagic balance possibly leading to the accumulation of damaged proteins and organelles contributing to disease progression. In cancer altered ATG7 expression may influence tumor survival by affecting cellular stress responses. Proteins such as p53 involved in cell cycle regulation often show association with ATG7-related pathways indicating a complex network influencing disease states. Understanding ATG7's role in these conditions can help explore potential therapeutic strategies.
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3 product images
Western blot - Human ATG7 knockout HeLa cell line (ab283307)
False colour image of Western blot: Anti-ATG7 antibody [EPR6251] staining at 1/10000 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 Anti-ATG7 antibody [EPR6251] ab133528 was shown to bind specifically to ATG7. A band was observed at 75 kDa in wild-type HeLa cell lysates with no signal observed at this size in ATG7 knockout cell line ab283307 (knockout cell lysate Human ATG7 knockout HeLa cell lysate ab287353). To generate this image wild-type and ATG7 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 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °. 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: Western blot - Anti-ATG7 antibody [EPR6251] (Anti-ATG7 antibody [EPR6251] ab133528) at 1/10000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: ATG7 knockout HeLa cell lysate at 20 µg
Lane 3: HepG2 cell lysate at 20 µg
Lane 4: Jurkat cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 77 kDa
Observed band size: 75 kDa
Western blot - Human ATG7 knockout HeLa cell line (ab283307)
False colour image of Western blot: Anti-ATG7 antibody [EP1759Y] staining at 1/100000 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 Anti-ATG7 antibody [EP1759Y] ab52472 was shown to bind specifically to ATG7. A band was observed at 75 kDa in wild-type HeLa cell lysates with no signal observed at this size in ATG7 knockout cell line ab283307 (knockout cell lysate Human ATG7 knockout HeLa cell lysate ab287353). To generate this image wild-type and ATG7 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 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °. 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: Western blot - Anti-ATG7 antibody [EP1759Y] (Anti-ATG7 antibody [EP1759Y] ab52472) at 1/100000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: ATG7 knockout HeLa cell lysate at 20 µg
Lane 3: HepG2 cell lysate at 20 µg
Lane 4: Jurkat cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 77 kDa
Observed band size: 75 kDa
Sanger Sequencing - Human ATG7 knockout HeLa cell line (ab283307)
41 bp deletion in exon 4
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