Human GPNMB/Osteoactivin ELISA Kit PicoKine®

Transmembrane glycoprotein NMB is a protein that in humans is encoded by the GPNMB gene. In osteoblast progenitor cells, GPNMB works as a positive regulator of osteoblast differentiation during later stages of matrix maturation and mineralization that is mediated at least in part by BMP-2 in a SMAD1 dependent manner to promote osteoblast differentiation. GPNMB can enhance the repairing process in bone fracture, demonstrated by its high expression during chondrogenesis (soft callus) and osteogenesis (hard callus) compared to the intact femurs that is why Osteoactivin (OA) could be a novel therapeutic agent used to treat generalized osteoporosis or localized osteopenia during fracture repair by stimulating bone growth and regeneration. Similarly, GPNMB expression increases during osteoclast differentiation and it is functionally implicated in this process, possibly by promoting the fusion of osteoclast progenitor cells.

Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 261 223 225 245 238 15.58 6.5% Sample 2 2107 1918 1797 1774 1899 131.96 6.9% Sample 3 5612 4864 5831 5547 5463 361.75 6.6% 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 1 261 223 225 245 238 15.58 6.5% Sample 1 261 223 225 245 238 15.58 6.5% Sample 2 2107 1918 1797 1774 1899 131.96 6.9% Sample 2 2107 1918 1797 1774 1899 131.96 6.9% Sample 3 5612 4864 5831 5547 5463 361.75 6.6% Sample 3 5612 4864 5831 5547 5463 361.75 6.6%

Up-regulated in various cancer cells, including in glioblastoma multiforme. Expressed in many melanoma cells, as well as in tissue macrophages, including liver Kuppfer cells and lung alveolar macrophages, in podocytes and in some cells of the ciliary body of the eye (at protein level) . Hardly detectable in healthy brain. .

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 261 2107 5612 260 2287 5804 Standard deviation 12.26 124.31 432.12 12.48 169.23 539.77 CV (%) 4.7% 7.7% 7.7% 4.8% 7.4% 9.3% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 261 2107 5612 260 2287 5804 Standard deviation 12.26 124.31 432.12 12.48 169.23 539.77 CV (%) 4.7% 7.7% 7.7% 4.8% 7.4% 9.3% 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) 261 2107 5612 260 2287 5804 Mean (pg/ml) 261 2107 5612 260 2287 5804 Standard deviation 12.26 124.31 432.12 12.48 169.23 539.77 Standard deviation 12.26 124.31 432.12 12.48 169.23 539.77 CV (%) 4.7% 7.7% 7.7% 4.8% 7.4% 9.3% CV (%) 4.7% 7.7% 7.7% 4.8% 7.4% 9.3%

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.)

Cell membrane; Single-pass type I membrane protein. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. Detected at the cell surface in different types of cancers cells, including glioblastoma multiforme cells and most melanoma cell lines.

Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.023 0.08 0.126 0.226 0.424 0.749 1.388 2.093 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.023 0.08 0.126 0.226 0.424 0.749 1.388 2.093 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.023 0.08 0.126 0.226 0.424 0.749 1.388 2.093 O.D. 0.023 0.08 0.126 0.226 0.424 0.749 1.388 2.093

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.