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HTRF Human MDA5 Aggregation Kit, 500 Assay Points

The MDA5 aggregation kit is designed for the fast detection of MDA5 aggregates in cell supernatants.

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Feature	Specification
Application	Protein Quantification
Sample Volume	16 µL

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The MDA5 aggregation kit is designed for the fast detection of MDA5 aggregates in cell supernatants.

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Product Variants

Part #:

63ADK111PEG

List Price

USD 1,671.37

Your online price:

Part #:

63ADK111PEH

List Price

USD 20,558.66

Your online price:

For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption, and disposal requirements under European REACH regulations (EC 1907/2006).

Product information

Overview

MDA-5 is an RIG-I-like receptor that acts as a cytoplasmic sensor of double stranded RNA, 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 proinflammatory cytokines.

It is understood to form aggregates along RNA strands when performing its nucleic acid sensing functions.

It may also play an important role in enhancing natural killer cell function, and may be involved in growth inhibition and apoptosis in several tumor cell lines.

Specifications

Other Specifications

Application	Protein Quantification
Brand	HTRF
Detection Modality	HTRF
Product Group	Kit
Sample Volume	16 µL
Shipping Conditions	Shipped in Dry Ice
Target Class	Biomarkers
Target Species	Human
Technology	TR-FRET
Therapeutic Area	Infectious Diseases Inflammation
Unit Size	500 assay points

Video gallery

How it works

MDA5 aggregation assay principle

MDA5 aggregates are measured using a sandwich immunoassay involving an anti-MDA5 monoclonal antibody specific for MDA5 aggregates, and labelled with terbium-Cryptate or d2, ensuring assay quality, reproducibility, and signal quality. The specific HTRF signal generated is proportional to the amount of MDA5 aggregates.

1biomarkers-how-it-works-assay-principle-mda5-aggregation-63adk111peg-63adk111peh.svg

MDA5 aggregation assay protocol

The cell samples are transferred into the assay plate for the detection of MDA5 aggregates. The antibodies labelled with HTRF fluorophores may be pre-mixed and added in a single dispensing step to further streamline the assay procedure. The assay detection can be run in 96- to 384-well plates by simply resizing each addition volume proportionally

2biomarkers-how-it-works-assay-protocol-mda5-aggregation-63adk111peg-63adk111peh.svg

Assay validation

MDA5 aggregation in HUTU80 cells stimulated with Poly I;C

HUTU80 cells were plated in 96-well plates in complete culture medium, and incubated at 37°C, 5% CO2. The next day, cells were treated with Poly I:C (HMW)/LyoVec in low serum medium (0.5% FBS) for 2h at 37°C, 5% CO2. After treatment, cells were then lysed with supplemented lysis buffer #3 for 30 minutes at RT under gentle shaking, and 16 µL of lysate were transferred twice over into a low volume white microplate before the addition of 4 µL of the premixed HTRF aggregation MDA5 or Total-MDA5 detection reagents. The HTRF signal was recorded after ON incubation.