Human TREX1 knockout A549 cell line
- Advanced Validation
- What is this?
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TREX1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 1 and 4 bp insertion in exon 1. 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
3' 5' exonuclease TREX1, 3' repair exonuclease 1, AGS1, AGS5, CRV, DKFZp434J0310, DNase III, DRN3, Deoxyribonuclease III, dnaQ/mutD (E. coli) like, HERNS, Three prime repair exonuclease 1
- WB
Lab
Western blot - Human TREX1 knockout A549 cell line (AB266927)
Lanes 1-4 : Merged signal (red and green). Green - ab185228 observed at 34 kDa. Red - loading control ab7291 observed at 50 kDa.
ab185228 Anti-TREX1 antibody [EPR14985] was shown to specifically react with TREX1 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab266927 (knockout cell lysate ab257764) was used. Wild-type and TREX1 knockout samples were subjected to SDS-PAGE. ab185228 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-TREX1 antibody [EPR14985] (<a href='/en-us/products/primary-antibodies/trex1-antibody-epr14985-ab185228'>ab185228</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
Western blot - Human TREX1 knockout A549 cell line (ab266927)
Lane 2:
TREX1 knockout A549 cell lysate at 20 µg
Lane 3:
Raji cell lysate at 20 µg
Lane 4:
Daudi 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: 39 kDa
Observed band size: 34 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human TREX1 knockout A549 cell line (AB266927)
Allele-1 : 1 bp insertion in exon1
- Cell Culture
Unknown
Cell Culture - Human TREX1 knockout A549 cell line (AB266927)
Representative images of TREX1 knockout A549 cells, low and high confluency examples (top left and right respectively) and wild-type A549 cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using an EVOS M5000 microscope.
- Sanger seq
Unknown
Sanger Sequencing - Human TREX1 knockout A549 cell line (AB266927)
Allele-2 : 4 bp insertion in exon 1.
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, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and 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 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.
- Do not allow the cell density to exceed 7x104 cells/cm2.
Culture medium
F-12K + 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
TREX1 removes excessive DNA that might trigger immune responses. It is not a part of a large protein complex but works closely with substrates involved in DNA repair and replication. By degrading abnormal DNA TREX1 prevents the initiation of inappropriate immune responses that could result in autoimmunity. Its functionality is essential in preventing the cell from converting these DNA fragments into ligands for DNA sensors which could activate immune signaling pathways.
Pathways
TREX1 plays a significant role in the DNA damage response and innate immune pathways. In the DNA damage response pathway TREX1 functions alongside proteins like ATR and ATM which address DNA replication stress and repair. Within the innate immune pathway it interacts with cGAS (cyclic GMP-AMP synthase) which can become activated in response to cytosolic DNA leading to the production of type I interferons - powerful immune modulators.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Target data
Complementary Products
Primary Antibodies
AB185228
Anti-TREX1 antibody [EPR14985]
RabMAb|Recombinant|KO Validated|20ul selling size
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TREX1 antibody [EPR14985] (AB185228)](https://content.abcam.com/products/images/trex1-antibody-epr14985-ab185228--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img128875.jpg)
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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