DESCRIPTION

Dystrophin (DMD or BMD) is a large muscle protein whose mutations cause Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD), the childhood neuromuscular disorders that result in progressive muscle weakness, respiratory difficulties and cardiovascular dysfunction. Dystrophin is a crucial component of the dystrophin-glycoprotein complex which is essential for muscle membrane integrity and stability. Dystrophin is located on the cytoplasmic face of the sarcolemma and connects the cytoskeletal network to the sarcolemma and extracellular matrix.

PRINCIPLE OF THE ASSAY

An anti-tag antibody is pre-coated onto the bottom of wells. After adding antigen or samples, Tagged-Capture antibody and HRP-conjugated detection antibody, a sandwich complex is formed in the solution. TMB acts as a HRP substrate, and the solution color will change from colorless to blue. A stop solution containing sulfuric acid turns the solution yellow. The color intensity is proportional to the quantity of bound protein, which is measurable at 450 nm with the correction wavelength set at 630 nm.

PRECISION

Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested 8 times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in 16 separate assays to assess inter-assay precision.

Intra-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	8	1,311.9	24.5	1.9
2	8	331.1	7.6	2.3
3	8	163.3	5.6	3.4

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	16	1,411.3	47.8	3.4
2	16	340.7	12.9	3.8
3	16	171.4	6.6	3.9

RECOVERY

The recovery of human/mouse/rat Dystrophin spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type		Average% of Expected	Range (%)
Cell lysate	1:16	97	93-103
	1:32	94	90-98
Tissue homogenates	1:8	100	95-105
	1:16	103	100-105

LINEARITY

To assess the linearity of the assay, samples were diluted with the appropriate Sample Diluent to produce samples with values within the dynamic range of the assay. (The cell lysate was initially diluted 1:2. The tissue homogenates was initially diluted 1:4.)

		Cell lysate	Tissue homogenates
1:2	Average% of Expected	100	100
	Range (%)	-	-
1:4	Average% of Expected	91	96
	Range (%)	85-97	91-101
1:8	Average% of Expected	95	99
	Range (%)	82-107	93-106
1:16	Average% of Expected	85	105
	Range (%)	73-97	100-109

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human/mouse/rat Dystrophin produced at Sunlong Medical™.

Ordering & shipping

• Minimum order: 1 kit
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