Assay procedure was performed according to manufacturer instructions. However, standard curve (duplicates) was modified due to the low concentrations observed in fasted pigs. Standard concentrations were 100, 50, 25, 12.5, 6.25, 3.125 and 1.5625 microIU/mL, prepared by a 1:3 dilution of the first standard provided with the product, and consecutive 1:2 dilutions thereafter.
Plasma samples were added to each well by duplicate (100 microL) as suggested by the manufacturer instructions, with no dilution. All remaining steps were performed in accordance with manufacturer instrucitions, with no modificatons. Readings were performed at 450 nm.
Results were as follows:
Control animals: 6.63, 3.65, 1.10 and 4.67 microIU/mL (mean: 4.01±1.15 microIU/mL)
Treatment A: 54.95, 9.29, 8.77, 5.10 microIU/mL (mean: 19.53±11.84).
Mr. Antonio Rodriguez Sinovas
Submitted Oct 28 2014