the Producing Standard Curve The standard curve is a serial dilution consisting of known concentrations of testosterone (or other hormone). Your samples are compared to this curve to determine the unknown concentrations. Serially dilute the testosterone standard as follows: First label nine 13 X 100 test tubes #1 - #9. If you are running 1 or 2 plates use the following protocol: Add 1 ml of Assay Buffer 3 to tube #1 and 500 l of Assay Buffer 3 to tubes #2 - #9. Remove 40 l of buffer from tube #1. Add 40l of the testosterone standard supplied with the kit (50,000 pg/ml) to tube #1. Vortex thoroughly. Add 500 l of tube #1 to tube #2 and vortex thoroughly. Add 500 l of tube #2 to tube #3 and Continue vortex. this for tubes #4 - #9. These diluted standards should be used within 60 min of preparation.
The concentrations of testosterone in tubes #1 through #9 will be 2000, 1000, 500, 250, 125, 62.5, 31.25, 15.625, and 7.8125 pg/ml, respectively. Because you will add 100 l of each standard to duplicate wells on the EIA plate, the concentrations of the standards in the standard curve can also be expressed as ...