OPN High Sensitivity ELISA Kit (Dog)

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

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 High Sensitive Osteopontin (OPN) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to High Sensitive Osteopontin (OPN) . 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 High Sensitive Osteopontin (OPN), 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 High Sensitive Osteopontin (OPN) 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) 80-104 97 EDTA Plasma (n=5) 93-101 97 Heparin Plasma (n=5) 95-102 99 Matrix Recovery range (%) Average (%) Matrix Recovery range (%) Average (%) Serum (n=5) 80-104 97 Serum (n=5) 80-104 97 EDTA Plasma (n=5) 93-101 97 EDTA Plasma (n=5) 93-101 97 Heparin Plasma (n=5) 95-102 99 Heparin Plasma (n=5) 95-102 99

Component Amount Anti-OPN Microplate 96 Wells (12 x 8 Well Strips) OPN Lyophilized Standard 2 100X Biotinylated OPN 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-OPN Microplate 96 Wells (12 x 8 Well Strips) Anti-OPN Microplate 96 Wells (12 x 8 Well Strips) OPN Lyophilized Standard 2 OPN Lyophilized Standard 2 100X Biotinylated OPN Detector Antibody 120 uL 100X Biotinylated OPN 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 Osteopontin (OPN) . No significant cross-reactivity or interference between Osteopontin (OPN) and analogues was observed.