Mouse OPN / Osteopontin ELISA Kit PicoKine®

Osteopontin (OPN) also called urinary stone protein, secreted phosphoprotein 1 (SPP1), bone sialoprotein, and early T lymphocyte activation 1 (ETA1) . Osteopontin is a phosphorylated glycoprotein secreted to the mineralizing extrOPNllular matrix by osteoblasts during bone development. It is believed to facilitate the attachment of osteoblasts and osteoclasts to the extrOPNllular matrix, allowing them to perform their respective functions during osteogenesis. Osteopontin is presumably involved in stone formation as stone matrix. The deduced protein sequence reveals a 317-amino acid protein (34,982 Da) containing a 16-amino acid hydrophobic signal sequence and a 33,352-Da protein destines to undergo extensive post-translational modifications before being secreted from the cell. The gene is located on human chromosome 4. The standard product used in this kit is recombinant mouse OPN, consisting of 284 amino acid sequence with the molecular mass of 31.5KDa. As a result of glycosylation, the molecular mass is 65KDa.

Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 306 280 315 325 306 16.71 5.4% Sample 2 1986 2147 1896 1939 1992 94.98 4.7% Sample 3 5401 5537 5315 5912 5541 228.21 4.1% Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 306 280 315 325 306 16.71 5.4% Sample 1 306 280 315 325 306 16.71 5.4% Sample 2 1986 2147 1896 1939 1992 94.98 4.7% Sample 2 1986 2147 1896 1939 1992 94.98 4.7% Sample 3 5401 5537 5315 5912 5541 228.21 4.1% Sample 3 5401 5537 5315 5912 5541 228.21 4.1%

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 306 1986 5401 308 2000 4959 Standard deviation 14.68 117.17 405.07 15.09 142 447.11 CV (%) 4.8% 7.5% 7.5% 4.9% 7.1% 9% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 306 1986 5401 308 2000 4959 Standard deviation 14.68 117.17 405.07 15.09 142 447.11 CV (%) 4.8% 7.5% 7.5% 4.9% 7.1% 9% Intra-Assay Precision Inter-Assay Precision Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 n 16 16 16 24 24 24 Mean (pg/ml) 306 1986 5401 308 2000 4959 Mean (pg/ml) 306 1986 5401 308 2000 4959 Standard deviation 14.68 117.17 405.07 15.09 142 447.11 Standard deviation 14.68 117.17 405.07 15.09 142 447.11 CV (%) 4.8% 7.5% 7.5% 4.9% 7.1% 9% CV (%) 4.8% 7.5% 7.5% 4.9% 7.1% 9%

Description|Quantity Pre-coated 96-well strip microplate|1 Standard|2 vials Biotinylated antibody (100x) |100ul Avidin-Biotin-Peroxidase Complex (100x) |100ul Sample Diluent|30ml Antibody Diluent|12ml Avidin-Biotin-Peroxidase Diluent|12ml Color Developing Reagent (TMB) |10ml Stop Solution|10ml Wash Buffer (25x) |20ml Adhesive plate sealers|4

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.025 0.065 0.103 0.178 0.314 0.572 1.122 1.85 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.025 0.065 0.103 0.178 0.314 0.572 1.122 1.85 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.025 0.065 0.103 0.178 0.314 0.572 1.122 1.85 O.D. 0.025 0.065 0.103 0.178 0.314 0.572 1.122 1.85

Microplate Reader capable of reading absorbance at 450nm. Incubator. Automated plate washer (optional) . Pipettes and pipette tips capable of precisely dispensing 0.5 μl through 1 ml volumes of aqueous solutions. Multichannel pipettes are recommended for large amount of samples. Deionized or distilled water. 500ml graduated cylinders. Test tubes for dilution.