Human DDX58 (RIG-I) knockout A549 cell line
- Advanced Validation
- What is this?
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DDX58 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 2 bp deletion in exon 2 and 5 bp deletion in exon 2.
View Alternative Names
DDX58_HUMAN, DEAD (Asp Glu Ala Asp) box polypeptide 58, DEAD (Asp Glu Ala Asp/His) box polypeptide, DEAD box protein 58, DEAD/H (Asp Glu Ala Asp/His) box polypeptide RIG1, DKFZp434J1111, DKFZp686N19181, FLJ13599, Probable ATP-dependent RNA helicase DDX58, RIG-I, RLR 1, RNA helicase, RNA helicase RIG I, Retinoic acid-inducible gene 1 protein, Retinoic acid-inducible gene I protein, Rig-1, SGMRT2
- WB
Lab
Western blot - Human DDX58 (RIG-I) knockout A549 cell line (AB267116)
Lanes 1 - 2 : Merged signal (red and green). Green - ab180675 observed at 107 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.
ab180675 was shown to react with DDX58 in A549 wild-type cells in western blot with loss of signal observed in DDX58 knockout cell line ab267116 (DDX58 knockout cell lysate ab257916). Wild-type and DDX58 knockout A549 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab180675 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-RIG-I/DDX58 antibody [EPR18629] (<a href='/en-us/products/primary-antibodies/rig-i-ddx58-antibody-epr18629-ab180675'>ab180675</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
DDX58 knockout A549 cell lysate at 20 µg
Lane 2:
Western blot - Human DDX58 (RIG-I) knockout A549 cell line (ab267116)
Predicted band size: 107 kDa
Observed band size: 107 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human DDX58 (RIG-I) knockout A549 cell line (AB267116)
Allele-2 : 2 bp deletion in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human DDX58 (RIG-I) knockout A549 cell line (AB267116)
Allele-1 : 5 bp deletion in exon2
Reactivity data
Product details
Recommended control: Human wild-type A549 cell line (ab255450). 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
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
RIG-I contributes significantly to the innate immune response. It acts to sense viral RNA and triggers the production of type I interferons and other pro-inflammatory cytokines. This protein functions as part of a complex that includes MAVS (mitochondrial antiviral signaling protein) and other signaling adapters. Upon activation RIG-I undergoes a conformational change leading to the exposure of its CARDs which interact with CARDs of MAVS facilitating downstream signaling to induce an antiviral state in host cells.
Pathways
RIG-I plays a central role in the RNA sensing pathway critical for antiviral immunity. This pathway involves several steps beginning with the recognition of viral RNA leading to the activation of interferon regulatory factors like IRF3 and IRF7 as well as nuclear factor kappa B (NF-κB). These factors then promote the expression of interferon-stimulated genes (ISGs). RIG-I also relates closely to the Jak-STAT signaling pathway which is activated by interferons and enhances the transcription of ISGs further amplifying the antiviral response.
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
AB180675
Anti-RIG-I/DDX58 antibody [EPR18629]
RabMAb|Recombinant|KO Validated
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Cell Lines & Lysates
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Primary Antibodies
AB290729
Anti-MAVS antibody [EPR26357-91]
BOND RX™ Validated|20ul selling size|RabMAb|Recombinant
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MAVS antibody [EPR26357-91] (AB290729)](https://content.abcam.com/products/images/mavs-antibody-epr26357-91-ab290729--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img400892.jpg)
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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