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Calculate the mean absorbance for each set of duplicate standards, controls and samples, and subtract the average zero standard optical density. Plot the standard curve on log-log graph paper or using Sigma plot software, with standard concentration on the x-axis and absorbance on the y-axis. Draw the best-fit straight line through the standard points. Typical Data: These standard curves are for demonstration only. A standard curve must be run with each assay. Assay Diluent C Rat CINC-3 concentration (pg/mL) 1 10 100 1000 10000 O D =4 50 n m 0.01 0.1 1 10 Assay Diluent B Rat CINC-3 concentration (pg/mL) 1 10 100 1000 O D =4 50 n m 0.01 0.1 1 10 Sensitivity: The minimum detectable dose of CINC-3 is typically less than 2 pg/mL. Recovery: Recovery was determined by spiking various levels of Rat CINC-3 into serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 88.29 76-101 Plasma 91.87 72-111 Cell culture media 83.48 75-89 Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 93.91 91.73 104.7 Range ( %) 86-102 83-107 95-113 1:4 Average % of Expected 75.26 74.70 79.28 Range ( %) 67-83 68-83 73-86 Reproducibility: Intra-Assay: CV<10 % Inter-Assay: CV<12 %