Human ATG12 knockout THP-1 cell line
- Advanced Validation
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ATG12 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. 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
APG12-like, APG12L, ATG12 autophagy related 12 homolog, ATG12 autophagy related 12 homolog (S. cerevisiae), ATG12_HUMAN, Apg12, Autophagy 12, Autophagy-related protein 12, FBR93, HAPG12, Ubiquitin-like protein ATG12
- WB
Lab
Western blot - Human ATG12 knockout THP-1 cell line (AB277831)
False colour image of Western blot : Anti-ATG12 antibody [EPR4800] staining at 1/1000 dilution shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution shown in red. In Western blot ab109491 was shown to bind specifically to ATG12. A band likely to be the unfunctional complex with ATG5 was observed at 52 kDa in wild-type THP-1 cell lysates with no signal observed at this size in Atg12 knockout cell line ab277831 (knockout cell lysate ab278183) - unconjugated functional form not observed at 15 kDa. To generate this image wild-type and Atg12 knockout THP-1 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-ATG12 antibody [EPR4800] (<a href='/en-us/products/primary-antibodies/atg12-antibody-epr4800-ab109491'>ab109491</a>) at 1/1000 dilution
Lane 1:
Wild-type THP-1 cell lysate at 20 µg
Lane 2:
ATG12 knockout THP-1 cell lysate at 20 µg
Lane 2:
Western blot - Human ATG12 knockout THP-1 cell line (ab277831)
Lane 3:
HCT 116 cell lysate at 20 µg
Lane 4:
HeLa cell lysate at 20 µg
Predicted band size: 15 kDa
Observed band size: 52 kDa
false
- WB
Lab
Western blot - Human ATG12 knockout THP-1 cell line (AB277831)
False colour image of Western blot : Anti-ATG12 antibody staining at 1/500 dilution shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution shown in red. In Western blot ab155589 was shown to bind specifically to ATG12. A band likely to be the unfunctional complex with ATG5 was observed at 52 kDa in wild-type THP-1 cell lysates with no signal observed at this size in Atg12 knockout cell line ab277831 (knockout cell lysate ab278183) - unconjugated functional form not observed at 15 kD. To generate this image wild-type and Atg12 knockout THP-1 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-ATG12 antibody (<a href='/en-us/products/primary-antibodies/atg12-antibody-ab155589'>ab155589</a>) at 1/500 dilution
Lane 1:
Wild-type THP-1 cell lysate at 20 µg
Lane 2:
ATG12 knockout THP-1 cell lysate at 20 µg
Lane 2:
Western blot - Human ATG12 knockout THP-1 cell line (ab277831)
Lane 3:
HCT 116 cell lysate at 20 µg
Lane 4:
HeLa cell lysate at 20 µg
Predicted band size: 15 kDa
Observed band size: 52 kDa
false
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
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. Based on cell count, seed cells in an appropriate cell culture flask at a density of 4x105 cells/mL. Seeding density is given as a guide only and should be scaled to align with individual lab schedules.
4. Incubate the culture at 37°C incubator with 5% CO2. Cultures should be monitored daily.
5. THP-1 cells recover slowly from cryopreservation and therefore may not be ready for subculture for a number of days. Cells should be left as much as possible over this time and only subcultured when the cell density reaches 8x105 cells/mL.Small amounts of fresh media can be added until cell number/viability improves.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- Cells should be seeded at 2x105 - 3x105 cells/mL and subcultured when they have reached 8x105 cells/mL.
- It is not recommended to allow the cell density to exceed 1x106 cells/mL.
Culture medium
RPMI + 10% FBS + 0.05 mM beta-mercaptoethanol
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
ATG12 is vital for the formation of autophagosomes which are double-membraned vesicles that sequester cellular components for degradation. It functions as part of a complex by binding to ATG5 and further associates with ATG16L1 to form a larger ATG12-ATG5-ATG16 complex. This complex localizes to the outer membrane of the growing autophagosome facilitating elongation and closure of the vesicle making it effective in the regulation of autophagy.
Pathways
ATG12's interaction with other proteins like ATG5 places it in the macroautophagy pathway an important process maintaining cellular homeostasis and turnover of cellular components. In addition to this ATG12 also contributes to cellular responses to stress conditions via this pathway. Furthermore it has connections with signaling pathways involving proteins such as LC3 which marks autophagosomal membranes and interacts with ATG12-conjugated complexes enhancing the autophagic flux.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Suspension
Gender
Male
Target data
Complementary Products
Cell Lines & Lysates
AB276116
Human CASP1 (Caspase-1) knockout THP-1 cell line
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