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<strong> Figure 1 WB Validation in the Brain of Mouse and Rat </strong><br>Loading: 15 μg of lysate Antibodies: MDA5, 4037,2 μ g/mL , 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 2 WB Validation in Daudi Cells</strong><br>Loading: 15 μg of lysate Antibodies: MDA5, 4037, 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.Lane A:1 μg/mLLane B: 2 μg/mLLane C: 4 μg/mL
<strong>Figure 3 Immunohistochemistry Validation of MDA5 in Human Lymph Node</strong><br> Immunohistochemical analysis of paraffin-embedded human lymph node tissue using anti-MDA5 antibody (4037) at 2.5 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
<strong>Figure 4 Immunohistochemistry Validation of MDA5 in Human Liver</strong><br> Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-MDA5 antibody (4037) at 2 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
<strong>Figure 5 Immunohistochemistry Validation of MDA5 in Mouse Brain</strong><br> Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-MDA5 antibody (4037) at 2 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
<strong>Figure 6 Immunohistochemistry Validation of MDA5 in Rat Brain</strong><br> Immunohistochemical analysis of paraffin-embedded rat brain tissue using anti-MDA5 antibody (4037) at 2 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
Figure 1 WB Validation in the Brain of Mouse and Rat
Loading: 15 μg of lysate Antibodies: MDA5, 4037,2 μ g/mL , 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat Anti-Rabbit IgG HRP conjugate at 1:10000 dilution.
Trial Size

MDA5 Antibody

CATALOG NUMBER: 4037

Clonality:
Polyclonal
Tested Applications:
ELISA, IHC-P, WB
Host Species:
Rabbit
Species Reactivity:
Human, Mouse, Rat
Conjugate:
Unconjugated
Specifications
Host Species:
Rabbit
Species Reactivity:
Human, Mouse, Rat
Immunogen:
Anti-MDA5 antibody (4037) was raised against a peptide corresponding to 16 amino acid synthetic peptide near the carboxy terminus of human MDA5.

The immunogen is located within the last 50 amino acids of MDA5.
Conjugate:
Unconjugated
Tested Applications:
ELISA, IHC-P, WB
Application Note:
WB: 1-4 μg/mL; IHC-P: 1-2.5 μg/mL.

Antibody validated: Western Blot in human, mouse and rat samples; Immunohistochemistry in human, mouse, and rat samples. All other applications and species not yet tested.
Positive Control 1:
Cat. No. 1403 - Mouse Brain Tissue Lysate
Positive Control 2:
Cat. No. 1463 - Rat Brain Tissue Lysate
Predicted Molecular Weight:
Predicted: 117 kD

Observed: 117 kD
Purification:
MDA5 Antibody is affinity chromatography purified via peptide column.
Clonality:
Polyclonal
Isotype:
IgG
physical-state:
Liquid
Buffer:
MDA5 Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Storage Conditions:
MDA5 antibody can be stored at 4°C for three months and -20°C, stable for up to one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.
Ncbi Official Symbol:
IFIH1
Additional Names:
MDA5 Antibody: Hlcd, MDA5, MDA-5, RLR-2, IDDM19, RH116, Interferon-induced helicase C domain-containing protein 1, Clinically amyopathic dermatomyositis autoantigen 140 kDa, CADM-140 autoantigen
Protein Accession Number:
NP_071451
Protein Gi Number:
27886568
Ncbi Gene Id Number:
64135
User Note:
Optimal dilutions for each application to be determined by the researcher.
Background:
MDA5 Antibody: The innate immune system detects viral infection by recognizing various viral components and triggers antiviral responses. Like the toll-like receptor 3 (TLR3), the melanoma differentiation-associated protein 5 (MDA5) recognizes double-stranded (ds) RNA, a molecular pattern associated with viral infection. MDA5, a member of the DEAD/DEAH-box RNA helicase family, consists of an amino-terminal caspase recruitment domain (CARD) and a carboxyl-terminal RNA helicase domain similar to that of the related protein RIG-1. When stimulated by dsRNA, MDA5 recruits the adaptor protein VISA and ultimately causes the activation of IRF-3 and NF-κB. MDA5 and RIG-1 recognize different types of dsRNA, with MDA5 recognizing poly (I:C). MDA5-null mice were highly susceptible to infection with picornaviruses, which possess such sequences, demonstrating the importance of MDA5 in innate immunity.
Background Reference 1:
Akira S, Uematsu S, and Takeuchi O. Pathogen recognition and innate immunity. Cell 2006; 124:783-801.
Background Reference 2:
Hiscott J, Nguyen T-LA, Arguello M, et al. Manipulation of the nuclear factor-kappaB pathway and the innate immune response by viruses. Oncogene 2006; 25:6844-67.
Background Reference 3:
Kang D, Gopalrishnan RV, Lin L, et al. Expression analysis and genomic characterization of human melanoma differentiation associated gene-5, mda-5: a novel type I interferon-responsive apoptosis-inducing gene. Oncogene 2004; 23:1789-800.
Background Reference 4:
Andrejeva J, Childs KS, Young DF, et al. The V proteins of the paramyxoviruses bind the IFN-inducible RNA helicase, mda-5, and inhibit its activation of the IFN-beta promoter. Proc. Natl. Acad. Sci. USA 2004; 101:17264-9.

FOR RESEARCH USE ONLY

For additional information, visit ProSci’s Terms & Conditions Page.

Disclaimer:
Optimal dilutions/concentrations should be determined by the end user. The information provided is a guideline for product use. This product is for research use only.

CATALOG NUMBER:

4037

List Size:
0.02 mg, 0.1 mg

List Price:

Price range: $99.00 through $445.00

Datasheet
Shipping Info

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$35 Dry Ice Shipment fee may be required on some items.

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