| MDA5

MDA5 (melanoma differentiation-associated protein 5) is a RIG-I-like receptor dsRNA helicase enzyme that is encoded by the IFIH1 gene. MDA5 is part of the RIG-I-like receptor (RLR) family, which also includes RIG-I and LGP2, and functions as a pattern recognition receptor capable of detecting viruses. It is generally believed that MDA5 recognizes double stranded RNA (dsRNA) over 2000nts in length, however it has been shown that whilst MDA5 can detect and bind to cytoplasmic dsRNA, it is also activated by a high molecular weight RNA complex composed of ssRNA and dsRNA. For many viruses, effective MDA5-mediated antiviral responses are dependent on functionally active LGP2.
 

| Typical data

As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, wash technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data are provided below for reference only.

 

Concentration (ng/mL)	20	10	5	2.5	1.25	0.62	0.31	0
OD	2.35	1.35	0.8	0.48	0.34	0.26	0.22	0.09
Corrected OD	2.26	1.26	0.71	0.39	0.25	0.17	0.13	-

 

| Repeatability

Intra-assay Precision (Precision within an assay)：CV% < 10%; Inter-assay Precision (Precision between assays)：CV% < 10%
 

| Recovery

The recovery of Human MDA5 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

 

Sample Type	Range(%)	Average Recovery(%)
Tissue Homogenates(n=8)	92-105	102
Cell Lysates(n=8)	96-108	105

 

| Linearity

Samples were spiked with high concentrations of Human MDA5 and diluted with Universal Diluent to produce samples with values within the range of the assay.

 

		Tissue Homogenates(n=4)	Cell Lysates(n=4)
1 : 2	Range (%)	88-96	90-110
	Average (%)	92	96
1 : 4	Range (%)	87-108	105-115
	Average (%)	98	108