Recombinant Anti-RIG-I/DDX58 antibody [EPR18629] (ab180675)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18629] to RIG-I/DDX58
- Suitable for: WB, IP
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-RIG-I/DDX58 antibody [EPR18629]
See all RIG-I/DDX58 primary antibodies -
Description
Rabbit monoclonal [EPR18629] to RIG-I/DDX58 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IPmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: A549, 293, HeLa and Jurkat whole cell lysates; Human fetal kidney and stomach lysates. IP: Jurkat whole cell lysate.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR18629 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab180675 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB | (1) |
1/1000. Detects a band of approximately 107 kDa (predicted molecular weight: 107 kDa).
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IP |
1/100.
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Notes |
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WB
1/1000. Detects a band of approximately 107 kDa (predicted molecular weight: 107 kDa). |
IP
1/100. |
Target
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Function
Involved in innate immune defense against viruses. Upon interaction with intracellular dsRNA produced during viral replication, triggers a transduction cascade involving MAVS/IPS1, which results in the activation of NF-kappa-B, IRF3 and IRF7 and the induction of the expression of antiviral cytokines such as IFN-beta and RANTES (CCL5). Detects dsRNA produced from non-self dsDNA by RNA polymerase III, such as Epstein-Barr virus-encoded RNAs (EBERs). Essential for the production of interferons in response to RNA viruses including paramyxoviruses, influenza viruses, Japanese encephalitis virus and HCV. -
Tissue specificity
Present in vascular smooth cells (at protein level). -
Sequence similarities
Belongs to the helicase family.
Contains 2 CARD domains.
Contains 1 helicase ATP-binding domain.
Contains 1 helicase C-terminal domain. -
Domain
The repressor domain controls homomultimerization and interaction with MAVS.
The helicase domain is responsible for dsRNA recognition.
The 2 CARD domains are responsible for interaction with and signaling through MAVS.
The second CARD domain is the primary site for 'Lys-63'-linked ubiquitination. -
Post-translational
modificationsIsgylated. Conjugated to ubiquitin-like protein ISG15 upon IFN-beta stimulation.
Ubiquitinated. Undergoes 'Lys-63'-linked ubiquitination. Lys-172 is the critical site for TRIM25-mediated ubiquitination, for MAVS binding and to induce anti-viral signal transduction. Lys-154, Lys-164 and Lys-172 are critical sites for RNF135-mediated ubiquitination. Deubiquitinated by CYLD, a protease that selectively cleaves 'Lys-63'-linked ubiquitin chains. -
Cellular localization
Cytoplasm. Colocalized with TRIM25 at cytoplasmic perinuclear bodies. - Information by UniProt
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Database links
- Entrez Gene: 23586 Human
- Omim: 609631 Human
- SwissProt: O95786 Human
- Unigene: 190622 Human
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Alternative names
- Ddx58 antibody
- DDX58_HUMAN antibody
- DEAD (Asp Glu Ala Asp) box polypeptide 58 antibody
see all
Images
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All lanes : Anti-RIG-I/DDX58 antibody [EPR18629] (ab180675) at 1/1000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : DDX58 knockout A549 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 107 kDa
Observed band size: 107 kDaLanes 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 ab267117 (DDX58 knockout cell lysate ab257917). 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.
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All lanes : Anti-RIG-I/DDX58 antibody [EPR18629] (ab180675) at 1/1000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : DDX58 knockout A549 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 107 kDa
Observed band size: 107 kDaLanes 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 wild-type A549 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.
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: RIG-I/DDX58 knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: K562 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab180675 observed at 107 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab180675 was shown to specifically react with RIG-I/DDX58 when RIG-I/DDX58 knockout samples were used. Wild-type and RIG-I/DDX58 knockout samples were subjected to SDS-PAGE. ab180675 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10 000 respectively and incubated overnight at 4°C. 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 1/10 000 dilution for 1 h at room temperature before imaging. -
All lanes : Anti-RIG-I/DDX58 antibody [EPR18629] (ab180675) at 1/1000 dilution
Lane 1 : 293 (Human epithelial cells from embryonic kidney) whole cell lysate
Lane 2 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Predicted band size: 107 kDa
Observed band size: 107 kDa
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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Anti-RIG-I/DDX58 antibody [EPR18629] (ab180675) at 1/1000 dilution + Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Predicted band size: 107 kDa
Observed band size: 107 kDa
Exposure time: 1 minuteBlocking/Dilution buffer: 5% NFDM/TBST.
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Anti-RIG-I/DDX58 antibody [EPR18629] (ab180675) at 1/1000 dilution + Human fetal kidney lysate at 10 µg
Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution
Predicted band size: 107 kDa
Observed band size: 107 kDa
Exposure time: 1 minuteBlocking/Dilution buffer: 5% NFDM/TBST.
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Anti-RIG-I/DDX58 antibody [EPR18629] (ab180675) at 1/1000 dilution + Human stomach lysate at 10 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Predicted band size: 107 kDa
Observed band size: 107 kDa
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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RIG-I/DDX58 was immunoprecipitated from 1mg of Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate with ab180675 at 1/100 dilution. Western blot was performed from the immunoprecipitate using ab180675 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: Jurkat whole cell lysate 10ug (Input). Lane 2: ab180675 IP in Jurkat whole cell lysate. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab180675 in Jurkat whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (11)
ab180675 has been referenced in 11 publications.
- Liu Y et al. RIG-I Promotes Cell Viability, Colony Formation, and Glucose Metabolism and Inhibits Cell Apoptosis in Colorectal Cancer by NF-κB Signaling Pathway. Dis Markers 2022:1247007 (2022). PubMed: 35242239
- You J et al. Hepatic exosomes with declined MiR-27b-3p trigger RIG-I/TBK1 signal pathway in macrophages. Liver Int 42:1676-1691 (2022). PubMed: 35460174
- Lei L et al. Comprehensive Diagnostics of Diabetic Nephropathy by Transcriptome RNA Sequencing. Diabetes Metab Syndr Obes 15:3069-3080 (2022). PubMed: 36237968
- Dunker W et al. TDP-43 prevents endogenous RNAs from triggering a lethal RIG-I-dependent interferon response. Cell Rep 35:108976 (2021). PubMed: 33852834
- Werner A et al. Widespread formation of double-stranded RNAs in testis. Genome Res 31:1174-86 (2021). PubMed: 34158368