Human OAS1 knockout A549 cell line
- Advanced Validation
- What is this?
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OAS1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 10 bp deletion in exon 2 and 1 bp insertion in exon 2. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View Alternative Names
(2 5')oligo(A) synthetase 1, (2-5'')oligo(A) synthase 1, 2 5 Oligoadenylate Synthetase 1, 2 5' oligo A synthase 1, 2 5' oligo A synthetase 1, 2 5A synthetase 1, 2' 5' oligo A synthetase 1, 2' 5' oligoadenylate synthetase 1, 2' 5' oligoadenylate synthetase 1 40/46kDa, 2' 5' oligoisoadenylate synthetase 1, 2''-5''-oligoadenylate synthase 1, 2-5A synthase 1, E18/E16, OAS1_HUMAN, OIAS, OIASI, p46/p42 OAS
- WB
Lab
Western blot - Human OAS1 knockout A549 cell line (AB267107)
False colour image of Western blot : Anti-OAS1 antibody 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, the antibody was shown to bind specifically to OAS1. A band was observed at 40 kDa in treated wild-type A549 cell lysates with no signal observed at this size in OAS1 knockout cell line ab267107 (knockout cell lysate ab259010). To generate this image, wild-type and OAS1 knockout A549 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 °C. 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:
Anti-OAS1 antibody at 1/1000 dilution
Lane 1:
Wild-type A549 Treated IFN-beta (1000 U/mL, 28 h) cell lysate at 20 µg
Lane 2:
OAS1 knockout A549 Treated IFN-beta (1000 U/mL, 28 h) cell lysate at 20 µg
Lane 2:
Western blot - Human OAS1 knockout A549 cell line (ab267107)
Lane 3:
Wild-type A549 control for IFN-beta (0 U/mL, 28 h) cell lysate at 20 µg
Lane 4:
OAS1 knockout A549 control for IFN-beta (0 U/mL, 28 h) cell lysate at 20 µg
Secondary
All lanes:
Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution
false
- Sanger seq
Unknown
Sanger Sequencing - Human OAS1 knockout A549 cell line (AB267107)
Allele-2 : 1 bp insertion in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human OAS1 knockout A549 cell line (AB267107)
Allele-1 : 10 bp deletion in exon2
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
The creation of 2'-5'-linked oligoadenylates by OAS1 is essential in activating RNase L an enzyme that degrades viral and cellular RNA to impede viral replication. OAS1 does not operate as part of a larger protein complex but functions independently to mediate antiviral activities. It has been observed that upon viral infection the levels of OAS1 increase enhancing the interferon response and cytokine signaling.
Pathways
OAS1 plays a significant role in the interferon signaling pathway and innate immune response pathways. Within these pathways OAS1 interacts with other proteins such as RNase L and is also associated with STAT1 a critical mediator of the cellular response to interferons. The function of OAS1 within these pathways highlights its importance in modulating immune responses to pathogens.
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
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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