Human DKK-3 PicoKine® Quick ELISA Kit

Dickkopf-related protein 3 is a protein that in humans is encoded by the DKK3 gene. This gene encodes a protein that is a member of the dickkopf family. It is mapped to 11p15.3. The secreted protein contains two cysteine rich regions and is involved in embryonic development through its interactions with the Wnt signaling pathway. The expression of this gene is decreased in a variety of cancer cell lines and it may function as a tumor suppressor gene. Members of the Dkk-related family display unique patterns of mRNA expression in human and mouse tissues, and are secreted when expressed in 293T cells. DKKs play an important role in vertebrate development, where they locally inhibit Wnt regulated processes such as antero-posterior axial patterning, limb development, somitogenesis and eye formation. In the adult, Dkks are implicated in bone formation and bone disease, cancer and Alzheimer disease.

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 1 326 288 312 330 314 23.55 7.5% Sample 2 1024 1226 1167 1048 1116 75.91 6.8% Sample 3 4848 4796 4999 4982 4906 240.41 4.9% 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 1 326 288 312 330 314 23.55 7.5% Sample 1 326 288 312 330 314 23.55 7.5% Sample 2 1024 1226 1167 1048 1116 75.91 6.8% Sample 2 1024 1226 1167 1048 1116 75.91 6.8% Sample 3 4848 4796 4999 4982 4906 240.41 4.9% Sample 3 4848 4796 4999 4982 4906 240.41 4.9%

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 326 1024 4848 254 1248 4526 Standard deviation 22.49 46.08 310.27 14.73 67.39 384.71 CV (%) 6.9% 4.5% 6.4% 5.8% 5.4% 8.5% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 326 1024 4848 254 1248 4526 Standard deviation 22.49 46.08 310.27 14.73 67.39 384.71 CV (%) 6.9% 4.5% 6.4% 5.8% 5.4% 8.5% 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) 326 1024 4848 254 1248 4526 Mean (pg/ml) 326 1024 4848 254 1248 4526 Standard deviation 22.49 46.08 310.27 14.73 67.39 384.71 Standard deviation 22.49 46.08 310.27 14.73 67.39 384.71 CV (%) 6.9% 4.5% 6.4% 5.8% 5.4% 8.5% CV (%) 6.9% 4.5% 6.4% 5.8% 5.4% 8.5%

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) 0 156 312 625 1,250 2,500 5,000 10,000 O.D. 0.071 0.240 0.403 0.637 1.110 1.782 2.153 2.214 Concentration (pg/ml) 0 156 312 625 1,250 2,500 5,000 10,000 O.D. 0.071 0.240 0.403 0.637 1.110 1.782 2.153 2.214 Concentration (pg/ml) 0 156 312 625 1,250 2,500 5,000 10,000 Concentration (pg/ml) 0 156 312 625 1,250 2,500 5,000 10,000 O.D. 0.071 0.240 0.403 0.637 1.110 1.782 2.153 2.214 O.D. 0.071 0.240 0.403 0.637 1.110 1.782 2.153 2.214

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.