AB240230

Anti-RIG-I/DDX58 antibody [EPR18629] - BSA and Azide free

RabMAb
Recombinant
KO Validated
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Rabbit Recombinant Monoclonal RIG-I/DDX58 antibody. Carrier free. Suitable for IP, WB and reacts with Human samples.

View Alternative Names

DDX58, RIGI, Antiviral innate immune response receptor RIG-I, ATP-dependent RNA helicase DDX58, DEAD box protein 58, RIG-I-like receptor 1, RNA sensor RIG-I, Retinoic acid-inducible gene 1 protein, Retinoic acid-inducible gene I protein, RLR-1, RIG-1, RIG-I

3 Images

Supplier Data

Immunoprecipitation - Anti-RIG-I/DDX58 antibody [EPR18629] - BSA and Azide free (AB240230)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab180675).

All lanes:

Immunoprecipitation - Anti-RIG-I/DDX58 antibody [EPR18629] (<a href='/en-us/products/primary-antibodies/rig-i-ddx58-antibody-epr18629-ab180675'>ab180675</a>)

Predicted band size: 107 kDa

false

Lab

Western blot - Anti-RIG-I/DDX58 antibody [EPR18629] - BSA and Azide free (AB240230)

This data was developed using the same antibody clone in a different buffer formulation (ab180675).

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 (<a href='/en-us/products/cell-lines/human-ddx58-rig-i-knockout-a549-cell-line-ab267116'>ab267116</a>)

Predicted band size: 107 kDa

Observed band size: 107 kDa

false

Lab

Western blot - Anti-RIG-I/DDX58 antibody [EPR18629] - BSA and Azide free (AB240230)

This data was developed using the same antibody clone in a different buffer formulation (ab180675).

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 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.

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 (<a href='/en-us/products/cell-lines/human-ddx58-rig-i-knockout-a549-cell-line-ab267117'>ab267117</a>)

Predicted band size: 107 kDa

Observed band size: 107 kDa

false

Unconjugated

Anti-RIG-I/DDX58 antibody [EPR18629]

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR18629

Isotype

IgG

Carrier free

Yes

Reacts with

Human

Applications

IP, WB

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "IP-species-checked": "testedAndGuaranteed", "IP-species-dilution-info": "", "IP-species-notes": "<p></p>", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>" } } }

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Product details

ab240230 is the carrier-free version of ab180675.

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

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Properties and storage information

Form

Liquid

Purification technique

Affinity purification Protein A

Storage buffer

pH: 7.2 - 7.4 Constituents: PBS

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

+4°C

Storage information

Do Not Freeze

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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

RIG-I also known as DDX58 is an important protein with mass of approximately 102 kDa. This protein acts as a cytosolic sensor for viral double-stranded RNA playing an essential role in the detection of viral infections. It is expressed in many cell types including immune and epithelial cells. RIG-I consists of two N-terminal caspase activation and recruitment domains (CARDs) a DExD/H box helicase domain and a C-terminal regulatory domain. These domains enable RIG-I to recognize and bind viral RNA initiating downstream signaling for immune responses.

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.

RIG-I is often associated with viral infections such as hepatitis C and influenza. By detecting viral RNA RIG-I activates immune responses that help to control these infections. However dysfunction or aberrations in RIG-I signaling can lead to autoimmune disorders such as Aicardi-Goutières syndrome where there is an inappropriate response to self nucleic acids. In these disease contexts RIG-I interacts with MAVS and indirectly with proteins involved in regulating immune responses such as STING which plays a role in the innate immune response against DNA viruses.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:

Visit the General protocols
Visit the Troubleshooting

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Target data

Innate immune receptor that senses cytoplasmic viral nucleic acids and activates a downstream signaling cascade leading to the production of type I interferons and pro-inflammatory cytokines (PubMed : 15208624, PubMed : 15708988, PubMed : 16125763, PubMed : 16127453, PubMed : 16153868, PubMed : 17190814, PubMed : 18636086, PubMed : 19122199, PubMed : 19211564, PubMed : 24366338, PubMed : 28469175, PubMed : 29117565, PubMed : 31006531, PubMed : 34935440, PubMed : 35263596, PubMed : 36793726). Forms a ribonucleoprotein complex with viral RNAs on which it homooligomerizes to form filaments (PubMed : 15208624, PubMed : 15708988). The homooligomerization allows the recruitment of RNF135 an E3 ubiquitin-protein ligase that activates and amplifies the RIG-I-mediated antiviral signaling in an RNA length-dependent manner through ubiquitination-dependent and -independent mechanisms (PubMed : 28469175, PubMed : 31006531). Upon activation, associates with mitochondria antiviral signaling protein (MAVS/IPS1) that activates the IKK-related kinases TBK1 and IKBKE which in turn phosphorylate the interferon regulatory factors IRF3 and IRF7, activating transcription of antiviral immunological genes including the IFN-alpha and IFN-beta interferons (PubMed : 28469175, PubMed : 31006531). Ligands include 5'-triphosphorylated ssRNAs and dsRNAs but also short dsRNAs (<1 kb in length) (PubMed : 15208624, PubMed : 15708988, PubMed : 19576794, PubMed : 19609254, PubMed : 21742966). In addition to the 5'-triphosphate moiety, blunt-end base pairing at the 5'-end of the RNA is very essential (PubMed : 15208624, PubMed : 15708988, PubMed : 19576794, PubMed : 19609254, PubMed : 21742966). Overhangs at the non-triphosphorylated end of the dsRNA RNA have no major impact on its activity (PubMed : 15208624, PubMed : 15708988, PubMed : 19576794, PubMed : 19609254, PubMed : 21742966). A 3'overhang at the 5'triphosphate end decreases and any 5'overhang at the 5' triphosphate end abolishes its activity (PubMed : 15208624, PubMed : 15708988, PubMed : 19576794, PubMed : 19609254, PubMed : 21742966). Detects both positive and negative strand RNA viruses including members of the families Paramyxoviridae : Human respiratory syncytial virus and measles virus (MeV), Rhabdoviridae : vesicular stomatitis virus (VSV), Orthomyxoviridae : influenza A and B virus, Flaviviridae : Japanese encephalitis virus (JEV), hepatitis C virus (HCV), dengue virus (DENV) and west Nile virus (WNV) (PubMed : 21616437, PubMed : 21884169). It also detects rotaviruses and reoviruses (PubMed : 21616437, PubMed : 21884169). Detects and binds to SARS-CoV-2 RNAs which is inhibited by m6A RNA modifications (Ref.71). Also involved in antiviral signaling in response to viruses containing a dsDNA genome such as Epstein-Barr virus (EBV) (PubMed : 19631370). Detects dsRNA produced from non-self dsDNA by RNA polymerase III, such as Epstein-Barr virus-encoded RNAs (EBERs). May play important roles in granulocyte production and differentiation, bacterial phagocytosis and in the regulation of cell migration.

See full target information RIGI

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

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

Anti-RIG-I/DDX58 antibody [EPR18629] - BSA and Azide free

Immunoprecipitation - Anti-RIG-I/DDX58 antibody [EPR18629] - BSA and Azide free (AB240230)

Western blot - Anti-RIG-I/DDX58 antibody [EPR18629] - BSA and Azide free (AB240230)

Western blot - Anti-RIG-I/DDX58 antibody [EPR18629] - BSA and Azide free (AB240230)

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Key facts

Host species

Clonality

Clone number

Isotype

Carrier free

Reacts with

Applications

Immunogen

Reactivity data

Product details

Properties and storage information

Form

Purification technique

Storage buffer

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Storage information

Supplementary information

Biological function summary

Pathways

Product protocols

Target data

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