Human ATG3 knockout HEK-293T cell line
- Advanced Validation
- What is this?
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Human ATG3 knockout HEK-293T cell line available to order. Recommended control: Human wild-type HEK293T cell line (ab255449).
View Alternative Names
2610016C12Rik, APG3 autophagy 3 like, APG3, S. cerevisiae, homolog of, APG3-like, APG3L, ATG3 autophagy related 3 homolog, ATG3 autophagy related 3 homolog (S. cerevisiae), ATG3_HUMAN, Apg 3, Apg3p, Autophagy 3, S. cerevisiae, homolog of, Autophagy Apg3p/Aut1p like, Autophagy-related protein 3, DKFZp564M1178, FLJ22125, MGC15201, OTTHUMP00000214547, OTTHUMP00000214548, PC3 96, Protein PC3-96, Ubiquitin-like-conjugating enzyme ATG3, autophagy related 3, hApg3
- WB
Lab
Western blot - Human ATG3 knockout HEK-293T cell line (AB266707)
Lanes 1-4 : Merged signal (red and green). Green - ab108251 observed at 40 kDa. Red - loading control ab7291 observed at 50 kDa.
ab108251 Anti-ATG3 antibody [EPR4801] was shown to specifically react with ATG3 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266707 (knockout cell lysate ab257363) was used. Wild-type and ATG3 knockout samples were subjected to SDS-PAGE. ab108251 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4° at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-ATG3 antibody [EPR4801] (<a href='/en-us/products/primary-antibodies/atg3-antibody-epr4801-ab108251'>ab108251</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK293T cell lysate at 20 µg
Lane 2:
ATG3 knockout HEK293T cell lysate at 20 µg
Lane 2:
Western blot - Human ATG3 knockout HEK-293T cell line (ab266707)
Lane 3:
K-562 cell lysate at 20 µg
Lane 4:
HeLa cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 35 kDa
Observed band size: 40 kDa
false
- WB
Lab
Western blot - Human ATG3 knockout HEK-293T cell line (AB266707)
Lanes 1-4 : Merged signal (red and green). Green - ab108282 observed at 40 kDa. Red - loading control ab7291 observed at 50 kDa.
ab108282 Anti-ATG3 antibody [EPR4802] was shown to specifically react with ATG3 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266707 (knockout cell lysate ab257363) was used. Wild-type and ATG3 knockout samples were subjected to SDS-PAGE. ab108282 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-ATG3 antibody [EPR4802] (<a href='/en-us/products/primary-antibodies/atg3-antibody-epr4802-ab108282'>ab108282</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK293T cell lysate at 20 µg
Lane 2:
ATG3 knockout HEK293T cell lysate at 20 µg
Lane 2:
Western blot - Human ATG3 knockout HEK-293T cell line (ab266707)
Lane 3:
K-562 cell lysate at 20 µg
Lane 4:
HeLa cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 35 kDa
Observed band size: 40 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human ATG3 knockout HEK-293T cell line (AB266707)
Homozygous : 1 bp deletion in exon 1
Reactivity data
Product details
Recommended control: Human wild-type HEK293T cell line (ab255449). 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.
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
ATG3 plays an important role in the elongation of the autophagic membrane. It operates as a part of the autophagy machinery working closely with ATG7 an E1-like enzyme to transfer phosphatidylethanolamine (PE) to LC3 converting it into LC3-II. This lipidated form of LC3 is essential in the expansion and closure of the isolation membrane which forms the autophagosome the core structure for degradation and recycling of cellular components.
Pathways
ATG3 is a central player in the macroautophagy pathway a process critical for cellular homeostasis. It interacts with ATG7 to facilitate the lipidation of ATG8 family proteins which are important for autophagosome formation. Moreover ATG3 also links to apoptosis pathways where its balance with pro-apoptotic proteins helps regulate cell death 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
Product protocols
- Visit the General protocols
- Visit the Troubleshooting
Target data
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