Human ATG16L1 knockout THP-1 cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Lab
Western blot - Human ATG16L1 knockout THP-1 cell line (AB277834)
False colour image of Western blot : Anti-ATG16L1 antibody [EPR15638] - N-terminal 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 ab187671 was shown to bind specifically to ATG16L1. A band was observed at 68/70 kDa in wild-type THP-1 cell lysates with no signal observed at this size in ATG16L1 knockout cell line ab277834 (knockout cell lysate ab278184). To generate this image wild-type and ATG16L1 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-ATG16L1 antibody [EPR15638] - N-terminal (<a href='/en-us/products/primary-antibodies/atg16l1-antibody-epr15638-n-terminal-ab187671'>ab187671</a>) at 1/1000 dilution
Lane 1:
Wild-type THP-1 cell lysate at 20 µg
Lane 2:
Western blot - Human ATG16L1 knockout THP-1 cell lysate (<a href='/en-us/products/cell-lysates/human-atg16l1-knockout-thp-1-cell-lysate-ab278184'>ab278184</a>) at 20 µg
Lane 3:
Wild type HeLa cell lysate at 20 µg
Lane 4:
ATG16L1 knockout HeLa cell lysate at 20 µg
Predicted band size: 68 kDa
Observed band size: 68 kDa,70 kDa
false
- WB
Lab
Western blot - Human ATG16L1 knockout THP-1 cell line (AB277834)
False colour image of Western blot : Anti-ATG16L1 antibody [5H9A11] staining at 1/500 dilution shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution shown in red. In Western blot ab233796 was shown to bind specifically to ATG16L1. A band was observed at 68/70 kDa in wild-type THP-1 cell lysates with no signal observed at this size in ATG16L1 knockout cell line ab277834 (knockout cell lysate ab278184). To generate this image wild-type and ATG16L1 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 fluorescent western blot (TBS-based) blocking solution 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-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.
All lanes:
Western blot - Anti-ATG16L1 antibody [5H9A11] (<a href='/en-us/products/primary-antibodies/atg16l1-antibody-5h9a11-ab233796'>ab233796</a>) at 1/500 dilution
Lane 1:
Wild-type THP-1 cell lysate at 20 µg
Lane 2:
ATG16L1 knockout THP-1 cell lysate at 20 µg
Lane 2:
Western blot - Human ATG16L1 knockout THP-1 cell line (ab277834)
Lane 3:
Wild type HeLa cell lysate at 20 µg
Lane 4:
ATG16L1 knockout HeLa cell lysate at 20 µg
Predicted band size: 68 kDa
Observed band size: 68 kDa,70 kDa
false
Complementary Products
Cell Lines & Lysates
AB276116
Human CASP1 (Caspase-1) knockout THP-1 cell line
Advanced Validation
- 0
- 0
Primary Antibodies
AB187671
Anti-ATG16L1 antibody [EPR15638] - N-terminal
RabMAb|Recombinant|KO Validated|Trial Size|Trial Size|Trial Size|Trial Size
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG16L1 antibody [EPR15638] - N-terminal (AB187671)](https://content.abcam.com/products/images/atg16l1-antibody-epr15638-n-terminal-ab187671--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img84248.jpg)
- 5
- 5
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
ATG16L1 plays a role in mediating autophagy a vital cellular degradation process. It forms a complex with ATG5 and ATG12 necessary for the elongation of the autophagosome membrane. Apart from its role in autophagy ATG16L1 contributes to the regulation of innate immunity by influencing the secretion of inflammatory cytokines. Its presence is essential for maintaining cellular homeostasis and proper immune responses.
Pathways
ATG16L1 is important in pathways like autophagy and immunity. In the autophagy pathway it works alongside ATG5 and ATG12 to ensure the proper formation of autophagosomes which are structures that engulf and degrade unwanted cellular components. Furthermore in the immune response pathway it helps regulate inflammation by managing cytokine production and secretion showing interaction with proteins like NOD2.
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
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