DAG1 ELISA Kit (Human)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Dystrophin Associated Glycoprotein 1 (DAG1) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Dystrophin Associated Glycoprotein 1 (DAG1) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

156DAG; A3a; AGRNR; DAG; dystroglycan; dystroglycan 1 (dystrophin-associated glycoprotein 1) ; Dystrophin-associated glycoprotein 1; LGMDR16; MDDGA9; MDDGC7; MDDGC9.

The dystroglycan complex is involved in a number of processes including laminin and basement membrane assembly, sarcolemmal stability, cell survival, peripheral nerve myelination, nodal structure, cell migration, and epithelial polarization.

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Dystrophin Associated Glycoprotein 1 (DAG1) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Dystrophin Associated Glycoprotein 1 (DAG1) . Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Dystrophin Associated Glycoprotein 1 (DAG1), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm +/- 10nm. The concentration of Dystrophin Associated Glycoprotein 1 (DAG1) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Reconstitution & Storage Instructions Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Antibody Array (AA) Protocol

Matrix Recovery Range (%) Average (%) Serum (n=5) 78-98 92 EDTA Plasma (n=5) 85-99 90 Heparin Plasma (n=5) 80-93 86 Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) Serum (n=5) 78-98 92 Serum (n=5) 78-98 92 EDTA Plasma (n=5) 85-99 90 EDTA Plasma (n=5) 85-99 90 Heparin Plasma (n=5) 80-93 86 Heparin Plasma (n=5) 80-93 86

Component Amount Anti-DAG1 Microplate 96 Wells (12 x 8 Well Strips) DAG1 Lyophilized Standard 2 x 20 ng 100X Biotinylated DAG1 Detector Antibody 120 uL Avidin/HRP Conjugate 120 uL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Component Amount Component Amount Anti-DAG1 Microplate 96 Wells (12 x 8 Well Strips) Anti-DAG1 Microplate 96 Wells (12 x 8 Well Strips) DAG1 Lyophilized Standard 2 x 20 ng DAG1 Lyophilized Standard 2 x 20 ng 100X Biotinylated DAG1 Detector Antibody 120 uL 100X Biotinylated DAG1 Detector Antibody 120 uL Avidin/HRP Conjugate 120 uL Avidin/HRP Conjugate 120 uL Standard Diluent 1 x 20 mL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 9 mL TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Stop Solution 1 x 6 mL

This assay has high sensitivity and excellent specificity for detection of Dystrophin Associated Glycoprotein 1 (DAG1) . No significant cross-reactivity or interference between Dystrophin Associated Glycoprotein 1 (DAG1) and analogues was observed.