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  4. MDA5 Antibody (YA2399)

MDA5 Antibody (YA2399) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to MDA5.

For research use only. We do not sell to patients.

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Top Publications Citing Use of Products
  • WB: Western Blot;
  • IHC-P: Immunohistochemistry-Paraffin;
  • IHC-F: Immunohistochemistry-Frozen;
  • ICC/IF: Immunocytochemistry/Immunofluorescence;
  • IF-Tissue: Immunofluorescence-Tissue;
  • mIHC: Multiplex Immunohistochemical;
  • IP: Immunoprecipitation;
  • ChIP: Chromatin Immunoprecipitation;
  • FC: Flow Cytometry;
  • ELISA: Enzyme Linked Immunosorbent Assay
  • Product Detail

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Description

MDA5 Antibody (YA2399) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to MDA5.

Host

Rabbit

Clonality

Recombinant, Monoclonal

Molecular Weight
Predicted band size: 117 kDa;
Observed band size: 117 kDa
Note: Due to possible protein modifications or aggregation, the molecular weight should be confirmed by actual measurement, and the predicted value is for reference only.
Species Reactivity
Human
SwissProt ID
Gene ID
Immunogen

A synthesized peptide derived from human MDA5 aa950-1025.

Application &
Dilution Ratio
Application Dilution Ratio
WB
WB: Western Blot
1:500-1:1000
ICC/IF
ICC/IF: Immunocytochemistry/Immunofluorescence
1:50-1:200
Sensitivity Endogenous Purity Affinity Chromatography
Conjugation Non-conjugated Modification Unmodified
Isotype IgG  
Appearance

Liquid

Formulation

Supplied in 50mM Tris-Glycine(pH 7.4), 0.15M NaCl, 40%Glycerol, 0.01% sodium azide and 0.05% BSA.

Storage & Stability

Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

Shipping

Shipping with blue ice.

Verification Image
ALL WB ICC
  • Western blot analysis of extracts from THP-1(lane 2(20ug) ,Jurkat(lane 3(20ug) and HEK293(lane 4(20ug) using MDA5 Antibody (HY-P82654) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (Beta Actin, HY-P80438, 1/10000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (1/10000) was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of HepG2 cells labeling MDA5 with MDA5 Antibody (HY-P82654) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with MDA5 Antibody (HY-P82654) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L(HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).

  • Immunocytochemistry analysis of HepG2 cells labeling MDA5 with MDA5 Antibody (HY-P82654) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with MDA5 Antibody (HY-P82654) at 1/100 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L(HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).

Background
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
Subcellular Localization:Cytoplasm; Nucleus; Mitochondrion
Expression:
Tissue_specificity:Widely expressed, at a low level. Expression is detected at slightly highest levels in placenta, pancreas and spleen and at barely levels in detectable brain, testis and lung

Induction:By interferon (IFN) and TNF
Isoforms & Post-Translational Modification:Q9BYX4 has 2 isomers: Q9BYX4-1: 116689 Da (predicted); Q9BYX4-2: 25129 Da (predicted).
Sumoylated. Sumoylation positively regulates its role in type I interferon induction and is enhanced by PIAS2-beta;Ubiquitinated by RNF125, leading to its degradation by the proteasome (PubMed:17460044). USP17/UPS17L2-dependent deubiquitination positively regulates the receptor (PubMed:20368735). Ubiquitinated by TRIM25 via 'Lys-63'-linked ubiquitination, promoting activation of IFIH1/MDA5 (PubMed:30193849). Ubiquitinated by TRIM40 via 'Lys-48'-linked ubiquitination; leading to proteasomal degradation (PubMed:29117565). Ubiquitinated by TRIM65 via 'Lys-63'-linked ubiquitination, promoting activation of IFIH1/MDA5 (PubMed:28594402);ISGylated by ISG15. ISGylation increases upon infection with dengue (DENV) or Zika (ZIKV) viruses. ISGylation at Lys-23 and Lys-43 is dependent of dephosphorylation at Ser-88, regulates mitochondrial translocation and oligomerization. Essential for IFIH1/MDA5-mediated cytokine responses and restriction of virus replication;Phosphorylated at Ser-88 (PubMed:23499489, PubMed:33727702). Dephosphorylated by phosphatases PPP11CA/PPP11CC; dephosphorylation precedes and is required for ISGylation (PubMed:23499489, PubMed:33727702);During apoptosis, processed into 3 cleavage products. The helicase-containing fragment, once liberated from the CARD domains, translocate from the cytoplasm to the nucleus. The processed protein significantly sensitizes cells to DNA degradation;(Microbial infection) Cleaved and inactivated by the protease 2A of coxsackievirus B3, poliovirus and enterovirus 71 allowing the virus to disrupt the host type I interferon production
Subunit:Monomer in the absence of ligands and homodimerizes in the presence of dsRNA ligands. Can assemble into helical or linear polymeric filaments on long dsRNA (PubMed:33727702). Interacts with MAVS/IPS1. Interacts (via the CARD domains) with TKFC, the interaction is inhibited by viral infection (PubMed:17600090). Interacts with PCBP2. Interacts with NLRC5. Interacts with PIAS2-beta. Interacts with DDX60. Interacts with ANKRD17. Interacts with IKBKE (PubMed:17600090). Interacts with ATG5 and ATG12, either as ATG5 and ATG12 monomers or as ATG12-ATG5 conjugates (PubMed:17709747). Interacts with ZCCHC3; leading to activate IFIH1/MDA5 (PubMed:30193849). Interacts with RNF123 (PubMed:27312109). Interacts with DDX3X (PubMed:20127681). Interacts with NOD1; this interaction promotes transcription of antiviral genes and inhibition of viral replication (PubMed:32169843). Interacts with ECSIT; this interaction bridges IFIH1 to the MAVS complex at the mitochondrion (PubMed:25228397)
RRID
Database
Research Field

Immunology

Synonyms
Helicard; Hlcd; IDDM19; IFIH1; MDA-5; RH116; RIG I like receptor 2; RLR 2; RNA helicase DEAD box protein 116
Documentation
Help & FAQs
  • Do most proteins show cross-species activity?

    Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

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Product Name:
MDA5 Antibody (YA2399)
Cat. No.:
HY-P82654
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