Human ATG7 knockout HeLa cell line
- Advanced Validation
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ATG7 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, functional Homozygous: 41 dp deletion in exon 4. 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
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
- WB
Lab
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] (ab8245) loading control staining at 1/20000 dilution shown in red. In Western blot 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 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-ATG7 antibody [EPR6251] (<a href='/en-us/products/primary-antibodies/atg7-antibody-epr6251-ab133528'>ab133528</a>) 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 2:
Western blot - Human ATG7 knockout HeLa cell line (ab283307)
Lane 3:
HepG2 cell lysate at 20 µg
Lane 4:
Jurkat cell lysate at 20 µg
Predicted band size: 77 kDa
Observed band size: 75 kDa
false
- WB
Lab
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] (ab8245) loading control staining at 1/20000 dilution shown in red. In Western blot 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 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-ATG7 antibody [EP1759Y] (<a href='/en-us/products/primary-antibodies/atg7-antibody-ep1759y-ab52472'>ab52472</a>) 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 2:
Western blot - Human ATG7 knockout HeLa cell line (ab283307)
Lane 3:
HepG2 cell lysate at 20 µg
Lane 4:
Jurkat cell lysate at 20 µg
Predicted band size: 77 kDa
Observed band size: 75 kDa
false
- Sanger seq
Lab
Sanger Sequencing - Human ATG7 knockout HeLa cell line (AB283307)
41 bp deletion in exon 4
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 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.
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
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.
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
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Advanced Validation
- 0
- 0
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