Invitrogen

MDA5 Polyclonal Antibody

Advanced Verification

This Antibody was verified by Cell treatment to ensure that the antibody binds to the antigen stated.

View all (22) MDA5 antibodies

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Datasheet

Protocols

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MDA5 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of Interferon-induced helicase C domain-containing protein 1 was performed using 70% confluent log phase A549 IFN gamma, 100 units/mL for 24 hrs. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with MDA5 Polyclonal Antibody (Product # PA5-82028) at 5 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32723), (1:2000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing Cytoplasmic localization. Panel e represents no signal was observed Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

MDA5 Antibody in Immunohistochemistry (IHC) — Immunohistochemical analysis of MDA5 in human placenta using a MDA5 polyclonal antibody (Product # PA5-82028). The analysis shows moderate cytoplasmic positivity in trophoblastic cells.

MDA5 Antibody in Western Blot (WB) — Knockout of MDA5 was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRIPSR637731_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of MDA5 was performed by loading 50 µg of HeLa Cas9 (Lane 1), HeLa Cas9 treated with 100 units/mL IFN gamma for 24hrs (Lane 2), HeLa MDA5 KO (Lane 3), and HeLa MDA5 KO treated with 100 units/mL IFN gamma for 24hrs (Lane 4) whole cell extracts. The samples were electrophoresed using NuPAGE™ 3 to 8%, Tris-Acetate, 1.0 mm, Mini Protein Gel (Product # EC6695BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with Anti-MDA5 Polyclonal Antibody (Product # PA5-82028, 1:200 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:8000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to MDA5.

MDA5 Antibody — Antibody specificity was demonstrated by CRISPR-Cas9 mediated knockout of target protein. A loss of signal was observed for target protein inMDA5 KO cell line compared to control cell line using Anti-MDA5 Polyclonal Antibody (Product # PA5-82028). {KO}

Product Details

PA5-82028

Applications	Tested Dilution	Publications
Western Blot (WB)	0.04-0.4 µg/mL	-
Immunohistochemistry (Paraffin) (IHC (P))	1:50-1:200	-

Product Specifications
Species Reactivity	Human
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	Recombinant protein corresponding to Human IFIH1 View immunogen
Conjugate	Unconjugated
Form	Liquid
Concentration	0.19 mg/mL
Purification	Anitgen affinity chromatography
Storage buffer	PBS, pH 7.2, with 40% glycerol
Contains	0.02% sodium azide
Storage conditions	Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
RRID	AB_2789189

Product Specific Information

Immunogen sequence: TIRMIDAYTH LETFYNEEKD KKFAVIEDDS DEGGDDEYCD GDEDEDDLKK PLKLDETDRF LMTLFFENNK MLKRLAENPE YENEKLTKLR NTIMEQYTRT EESARGIIFT KTRQSAYALS QWITENEKFA EVGVKAHHLI GAGHSSEFKP

Target Information

DEAD box proteins, characterized by the conserved motif Asp-Glu-Ala-Asp and a protein kinase C-activating compound, mezerein. Irreversible reprogramming of melanomas can be achieved by treatment with both these agents; treatment with either agent alone only achieves reversible differentiation.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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Bioinformatics

Protein Aliases: CADM-140 autoantigen; Clinically amyopathic dermatomyositis autoantigen 140 kDa; DEAD/H (Asp-Glu-Ala-Asp/His) box polypeptide; Helicard; Helicase with 2 CARD domains; interferon induced, with helicase C domain 1; Interferon-induced helicase C domain-containing protein 1; Interferon-induced with helicase C domain protein 1; MDA-5; melanoma differentiation associated protein-5; melanoma differentiation-associated gene 5; Melanoma differentiation-associated protein 5; MGC133047; Murabutide down-regulated protein; RIG-I-like receptor 2; RLR-2; RNA helicase-DEAD box protein 116

Gene Aliases: AGS7; Hlcd; IDDM19; IFIH1; MDA-5; MDA5; RH116; RLR-2; SGMRT1

UniProt ID: (Human) Q9BYX4

Entrez Gene ID: (Human) 64135

Function(s)

detection of virus response to virus protein sumoylation negative regulation of type I interferon production positive regulation of interferon-alpha production positive regulation of interferon-beta production regulation of type III interferon production cytoplasmic pattern recognition receptor signaling pathway in response to virus MDA-5 signaling pathway innate immune response

Process(es)

DNA binding double-stranded RNA binding single-stranded RNA binding helicase activity protein binding ATP binding zinc ion binding ribonucleoprotein complex binding