Rabbit Recombinant Monoclonal MDA5 antibody. Carrier free. Suitable for WB, IP, Flow Cyt (Intra), ICC/IF and reacts with Mouse samples.
Images
![Western blot - Anti-MDA5 antibody [EPR28350-21] - BSA and Azide free (AB315243), expandable thumbnail](https://content.abcam.com/products/images/mda5-antibody-epr28350-21-bsa-and-azide-free-ab315243--western-blot-img424421.jpg/_jcr_content/renditions/webp-475.webp "Western blot - Anti-MDA5 antibody [EPR28350-21] - BSA and Azide free (AB315243)")
![Immunocytochemistry/ Immunofluorescence - Anti-MDA5 antibody [EPR28350-21] - BSA and Azide free (AB315243), expandable thumbnail](https://content.abcam.com/products/images/mda5-antibody-epr28350-21-bsa-and-azide-free-ab315243--immunocytochemistry-immunofluorescence-img424497.jpg/_jcr_content/renditions/webp-475.webp "Immunocytochemistry/ Immunofluorescence - Anti-MDA5 antibody [EPR28350-21] - BSA and Azide free (AB315243)")
![Western blot - Anti-MDA5 antibody [EPR28350-21] - BSA and Azide free (AB315243), expandable thumbnail](https://content.abcam.com/products/images/mda5-antibody-epr28350-21-bsa-and-azide-free-ab315243--western-blot-img424420.jpg/_jcr_content/renditions/webp-475.webp "Western blot - Anti-MDA5 antibody [EPR28350-21] - BSA and Azide free (AB315243)")
![Western blot - Anti-MDA5 antibody [EPR28350-21] - BSA and Azide free (AB315243), expandable thumbnail](https://content.abcam.com/products/images/mda5-antibody-epr28350-21-bsa-and-azide-free-ab315243--western-blot-img424419.jpg/_jcr_content/renditions/webp-475.webp "Western blot - Anti-MDA5 antibody [EPR28350-21] - BSA and Azide free (AB315243)")
![Flow Cytometry (Intracellular) - Anti-MDA5 antibody [EPR28350-21] - BSA and Azide free (AB315243), expandable thumbnail](https://content.abcam.com/products/images/mda5-antibody-epr28350-21-bsa-and-azide-free-ab315243--flow-cytometry-intracellular-img424509.jpg/_jcr_content/renditions/webp-475.webp "Flow Cytometry (Intracellular) - Anti-MDA5 antibody [EPR28350-21] - BSA and Azide free (AB315243)")
Key facts
- Isotype
- IgG
- Host species
- Rabbit
- Storage buffer
pH: 7.2 - 7.4
Constituents: 100% PBS- Form
- Liquid
- Clonality
- Monoclonal
Immunogen
- The exact immunogen used to generate this antibody is proprietary information.
Reactivity data
Select an application| WB | IP | Flow Cyt (Intra) | ICC/IF | IHC-P | |
|---|---|---|---|---|---|
| Human | Not recommended | Not recommended | Not recommended | Not recommended | Not recommended |
| Mouse | Tested | Tested | Tested | Tested | Not recommended |
| Rat | Not recommended | Not recommended | Not recommended | Not recommended | Not recommended |
WB
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Mouse | Dilution info - | Notes - |
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Rat, Human | Dilution info - | Notes - |
IP
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Mouse | Dilution info - | Notes - |
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Rat, Human | Dilution info - | Notes - |
Flow Cyt (Intra)
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Mouse | Dilution info - | Notes - |
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Human, Rat | Dilution info - | Notes - |
ICC/IF
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Mouse | Dilution info - | Notes - |
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Human, Rat | Dilution info - | Notes - |
IHC-P
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Human, Mouse, Rat | Dilution info - | Notes - |
Associated Products
Select an associated product type
1 product for Alternative Version
Target data
Function
Innate immune receptor which acts as a cytoplasmic sensor of viral nucleic acids and plays a major role in sensing viral infection and in the activation of a cascade of antiviral responses including the induction of type I interferons and pro-inflammatory cytokines (PubMed:28594402, PubMed:32169843, PubMed:33727702). Its ligands include mRNA lacking 2'-O-methylation at their 5' cap and long-dsRNA (>1 kb in length) (PubMed:22160685). Upon ligand binding it associates with mitochondria antiviral signaling protein (MAVS/IPS1) which activates the IKK-related kinases: TBK1 and IKBKE which phosphorylate interferon regulatory factors: IRF3 and IRF7 which in turn activate transcription of antiviral immunological genes, including interferons (IFNs); IFN-alpha and IFN-beta. Responsible for detecting the Picornaviridae family members such as encephalomyocarditis virus (EMCV), mengo encephalomyocarditis virus (ENMG), and rhinovirus (PubMed:28606988). Detects coronavirus SARS-CoV-2 (PubMed:33440148, PubMed:33514628). Can also detect other viruses such as dengue virus (DENV), west Nile virus (WNV), and reovirus. Also involved in antiviral signaling in response to viruses containing a dsDNA genome, such as vaccinia virus. Plays an important role in amplifying innate immune signaling through recognition of RNA metabolites that are produced during virus infection by ribonuclease L (RNase L). May play an important role in enhancing natural killer cell function and may be involved in growth inhibition and apoptosis in several tumor cell lines.
Alternative names
MDA5, RH116, IFIH1, Interferon-induced helicase C domain-containing protein 1, Clinically amyopathic dermatomyositis autoantigen 140 kDa, Helicase with 2 CARD domains, Interferon-induced with helicase C domain protein 1, Melanoma differentiation-associated protein 5, Murabutide down-regulated protein, RIG-I-like receptor 2, RNA helicase-DEAD box protein 116, CADM-140 autoantigen, Helicard, MDA-5, RLR-2
Recommended products
Rabbit Recombinant Monoclonal MDA5 antibody. Carrier free. Suitable for WB, IP, Flow Cyt (Intra), ICC/IF and reacts with Mouse samples.
Key facts
- Isotype
- IgG
- Host species
- Rabbit
- Storage buffer
pH: 7.2 - 7.4
Constituents: 100% PBS- Form
- Liquid
- Clonality
- Monoclonal
- Immunogens
- The exact immunogen used to generate this antibody is proprietary information.
- Carrier free
- Yes
- Clone number
- EPR28350-21
- Purification technique
- Affinity purification Protein A
- Concentration
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- +4°C
Notes
ab315243 is the carrier-free version of Anti-MDA5 antibody [EPR28350-21] ab315242.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
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.
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
MDA5 also known as melanoma differentiation-associated protein 5 or IFIH1 is a cytosolic sensor involved in the detection of viral RNA. It is a member of the RIG-I-like receptor family and possesses a molecular weight of approximately 116 kilodaltons. MDA5 protein is expressed across various tissues but shows higher expression in the immune system's cells including macrophages and dendritic cells. MDA5 recognizes long double-stranded RNA a common viral replication byproduct triggering immune responses.
- Biological function summary
In cellular defense mechanisms MDA5 plays an important role in initiating antiviral responses. It operates as part of a signaling complex that includes MAVS (mitochondrial antiviral-signaling protein) leading to the production of type I interferons and pro-inflammatory cytokines. These responses promote an antiviral state in host cells aiding in the containment and clearance of viral infections. MDA5 antibodies can be useful tools for studying these immune processes.
- Pathways
MDA5 interacts with antiviral signaling pathways such as the IFN signaling pathway and the RIG-I-like receptor signaling pathway. Upon recognizing viral RNA MDA-5 cooperates with proteins like RIG-I and MAVS to activate downstream signaling cascades that mediate immune responses. This collaboration enables efficient viral recognition and response limiting viral replication and spread.
- Associated diseases and disorders
MDA5 has associations with autoimmune diseases like systemic lupus erythematosus and certain types of dermatomyositis. These conditions often involve autoantibodies against MDA5 indicating a possible role in pathogenesis. In these diseases altered function or regulation of related proteins including IFN-induced proteins may contribute to chronic inflammation and autoimmunity. Understanding MDA5's involvement in these contexts is essential for developing therapeutic strategies.
Product promise
Tested
We have tested this species and application combination and it works. It is covered by our product promise.
Expected
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
Predicted
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
Not recommended
We do not recommend this combination. It is not covered by our product promise.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
6 product images
![Western blot - Anti-MDA5 antibody [EPR28350-21] - BSA and Azide free (ab315243), expandable thumbnail](https://content.abcam.com/products/images/mda5-antibody-epr28350-21-bsa-and-azide-free-ab315243--western-blot-img424421.jpg "Western blot - Anti-MDA5 antibody [EPR28350-21] - BSA and Azide free (ab315243)")
Western blot - Anti-MDA5 antibody [EPR28350-21] - BSA and Azide free (ab315243)
This data was developed using Anti-MDA5 antibody [EPR28350-21] ab315242, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Low expression: testis (PMID: 12015121)
Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.
The identity of the higher MW band at approximately 250 kDa is unknown.
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-MDA5 antibody [EPR28350-21] (Anti-MDA5 antibody [EPR28350-21] ab315242) at 1/1000 dilution
Lane 1: Mouse liver tissue lysate at 20 µg
Lane 2: Mouse testis tissue lysate at 20 µg
SecondaryAll lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 140 kDa, 90 kDa, 36 kDa
Exposure time: 180s
![Immunocytochemistry/ Immunofluorescence - Anti-MDA5 antibody [EPR28350-21] - BSA and Azide free (ab315243), expandable thumbnail](https://content.abcam.com/products/images/mda5-antibody-epr28350-21-bsa-and-azide-free-ab315243--immunocytochemistry-immunofluorescence-img424497.jpg "Immunocytochemistry/ Immunofluorescence - Anti-MDA5 antibody [EPR28350-21] - BSA and Azide free (ab315243)")
Immunocytochemistry/ Immunofluorescence - Anti-MDA5 antibody [EPR28350-21] - BSA and Azide free (ab315243)
This data was developed using Anti-MDA5 antibody [EPR28350-21] ab315242, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cells labelling MDA5 with Anti-MDA5 antibody [EPR28350-21] ab315242 at 1/100 (5.09 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).
Confocal image showing increased cytoplasmic staining in RAW 264.7 cells treated with IFNβ (2000 U/mL) for 24 hr. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).
Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.
![Western blot - Anti-MDA5 antibody [EPR28350-21] - BSA and Azide free (ab315243), expandable thumbnail](https://content.abcam.com/products/images/mda5-antibody-epr28350-21-bsa-and-azide-free-ab315243--western-blot-img424420.jpg "Western blot - Anti-MDA5 antibody [EPR28350-21] - BSA and Azide free (ab315243)")
Western blot - Anti-MDA5 antibody [EPR28350-21] - BSA and Azide free (ab315243)
This data was developed using Anti-MDA5 antibody [EPR28350-21] ab315242, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Low expression: brain (PMID: 12015121)
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 17267501, 24390337).
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-MDA5 antibody [EPR28350-21] (Anti-MDA5 antibody [EPR28350-21] ab315242) at 1/1000 dilution
Lane 1: Mouse spleen tissue lysate at 40 µg
Lane 2: Mouse colon tissue lysate at 40 µg
Lane 3: Mouse brain tissue lysate at 40 µg
SecondaryAll lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 140 kDa, 90 kDa, 36 kDa
Exposure time: 180s
![Western blot - Anti-MDA5 antibody [EPR28350-21] - BSA and Azide free (ab315243), expandable thumbnail](https://content.abcam.com/products/images/mda5-antibody-epr28350-21-bsa-and-azide-free-ab315243--western-blot-img424419.jpg "Western blot - Anti-MDA5 antibody [EPR28350-21] - BSA and Azide free (ab315243)")
Western blot - Anti-MDA5 antibody [EPR28350-21] - BSA and Azide free (ab315243)
This data was developed using Anti-MDA5 antibody [EPR28350-21] ab315242, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-MDA5 antibody [EPR28350-21] (Anti-MDA5 antibody [EPR28350-21] ab315242) at 1/1000 dilution
Lane 1: Untreated RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg
Lane 2: RAW264.7 treated with 2000U/ml IFN beta for 24 hours whole cell lysate at 20 µg
SecondaryAll lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 140 kDa, 36 kDa
Exposure time: 15s
![Flow Cytometry (Intracellular) - Anti-MDA5 antibody [EPR28350-21] - BSA and Azide free (ab315243), expandable thumbnail](https://content.abcam.com/products/images/mda5-antibody-epr28350-21-bsa-and-azide-free-ab315243--flow-cytometry-intracellular-img424509.jpg "Flow Cytometry (Intracellular) - Anti-MDA5 antibody [EPR28350-21] - BSA and Azide free (ab315243)")
Flow Cytometry (Intracellular) - Anti-MDA5 antibody [EPR28350-21] - BSA and Azide free (ab315243)
This data was developed using Anti-MDA5 antibody [EPR28350-21] ab315242, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 2000U/ml IFN beta for 24h (Red) / Untreated RAW 264.7 (Green) cells labelling MDA5 with Anti-MDA5 antibody [EPR28350-21] ab315242 at 1/500 dilution (0.1 ug, Red and Green) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).
Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.
![Immunoprecipitation - Anti-MDA5 antibody [EPR28350-21] - BSA and Azide free (ab315243), expandable thumbnail](https://content.abcam.com/products/images/mda5-antibody-epr28350-21-bsa-and-azide-free-ab315243--immunoprecipitation-img424447.jpg "Immunoprecipitation - Anti-MDA5 antibody [EPR28350-21] - BSA and Azide free (ab315243)")
Immunoprecipitation - Anti-MDA5 antibody [EPR28350-21] - BSA and Azide free (ab315243)
This data was developed using Anti-MDA5 antibody [EPR28350-21] ab315242, the same antibody clone in a different buffer formulation.
MDA5 was immunoprecipitated from 0.35 mg Mouse spleen whole cell lysate with Anti-MDA5 antibody [EPR28350-21] ab315242 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-MDA5 antibody [EPR28350-21] ab315242 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: Mouse spleen whole cell lysate
Lane 2: Anti-MDA5 antibody [EPR28350-21] ab315242 IP in Mouse spleen whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-MDA5 antibody [EPR28350-21] ab315242 in mouse spleen whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
All lanes: Immunoprecipitation - Anti-MDA5 antibody [EPR28350-21] (Anti-MDA5 antibody [EPR28350-21] ab315242) at 1/30 dilution
All lanes: Mouse spleen whole cell lysate
SecondaryAll lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Exposure time: 180s
![Immunoprecipitation - Anti-MDA5 antibody [EPR28350-21] - BSA and Azide free (ab315243), expandable thumbnail](https://content.abcam.com/products/images/mda5-antibody-epr28350-21-bsa-and-azide-free-ab315243--immunoprecipitation-img424447.jpg/_jcr_content/renditions/webp-475.webp "Immunoprecipitation - Anti-MDA5 antibody [EPR28350-21] - BSA and Azide free (ab315243)")
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