Human TRANCE/RANKL/TNFSF11 PicoKine® Quick ELISA Kit

Receptor activator of nuclear factor kappa-B ligand (RANKL), also known as tumor necrosis factor ligand superfamily member 11 (TNFSF11), is a protein that in humans is encoded by the TNFSF11 gene. This gene encodes a member of the tumor necrosis factor (TNF) cytokine family which is a ligand for osteoprotegerin and functions as a key factor for osteoclast differentiation and activation. This gene is mapped to chromosome13q14.11. Targeted disruption of the related gene in mice led to severe osteopetrosis and a lack of osteoclasts. The deficient mice exhibited defects in early differentiation of T and B lymphocytes, and failed to form lobulo-alveolar mammary structures during pregnancy. This gene may play an important role in enhanced bone-resorption in humoral hypercalcemia of malignancy.

Reproducibility To assay reproducibility, three samples with differing target protein concentrations were assayed using four different lots. Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 117 127 119 129 123 4.67 3.8% Sample 2 867 748 853 803 817 41.66 5.1% Sample 3 2244 2228 2257 2225 2238 33.57 1.5% 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 1 117 127 119 129 123 4.67 3.8% Sample 1 117 127 119 129 123 4.67 3.8% Sample 2 867 748 853 803 817 41.66 5.1% Sample 2 867 748 853 803 817 41.66 5.1% Sample 3 2244 2228 2257 2225 2238 33.57 1.5% Sample 3 2244 2228 2257 2225 2238 33.57 1.5%

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 117 867 2244 116 799 2281 Standard deviation 5.85 55.48 109.95 6.26 60.72 120.89 CV (%) 5.0% 6.4% 4.9% 5.4% 7.6% 5.3% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 117 867 2244 116 799 2281 Standard deviation 5.85 55.48 109.95 6.26 60.72 120.89 CV (%) 5.0% 6.4% 4.9% 5.4% 7.6% 5.3% 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) 117 867 2244 116 799 2281 Mean (pg/ml) 117 867 2244 116 799 2281 Standard deviation 5.85 55.48 109.95 6.26 60.72 120.89 Standard deviation 5.85 55.48 109.95 6.26 60.72 120.89 CV (%) 5.0% 6.4% 4.9% 5.4% 7.6% 5.3% CV (%) 5.0% 6.4% 4.9% 5.4% 7.6% 5.3%

Description|Quantity|Volume Anti-tag Pre-coated 96-well Strip Microplate|1|12 strips of 8 wells Standard||2 Antibody A|1|3ml Antibody B|1|3ml Sample Diluent|1|15ml TBS-T Wash Buffer (25x) |1|12ml Color Developing Reagent (TMB) |1|10ml Stop Solution|1|10ml Adhesive Plate Sealers|2|Piece

Concentration (pg/ml) 0 78 156 312 625 1,250 2,500 5,000 O.D. 0.034 0.139 0.151 0.262 0.453 0.783 1.331 2.026 Concentration (pg/ml) 0 78 156 312 625 1,250 2,500 5,000 O.D. 0.034 0.139 0.151 0.262 0.453 0.783 1.331 2.026 Concentration (pg/ml) 0 78 156 312 625 1,250 2,500 5,000 Concentration (pg/ml) 0 78 156 312 625 1,250 2,500 5,000 O.D. 0.034 0.139 0.151 0.262 0.453 0.783 1.331 2.026 O.D. 0.034 0.139 0.151 0.262 0.453 0.783 1.331 2.026

Microplate Reader capable of reading absorbance at 450nm. 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. Horizontal orbital microplate shaker capable of maintaining a speed of 500 rpm, amplitude 3mm.