Bio-Rad Human Metabolic and Hormone Assays - Manual (Page 15)

13
Dilution of Standard (1:3 Serial Dilution)
This preparation provides sufficient volume to run duplicate standard
dilution curves. Ensure that each new standard is mixed well by vortexing
before proceeding to the next dilution. Change tips between each dilution.
Note: The product data sheet in each kit lists the most concentrated
point on the standard curve (S1). Enter the values and units into
Bio-Plex Manager
™ software as instructed in the Read Plate section.
1. Label 9 polypropylene tubes S1 through S8 and Blank.
2. Transfer the reconstituted standard into the tube labeled S1.
3. Add the appropriate amount of the standard diluent into the labeled
tubes according to Figure 3 (this will be sufficient for duplicate
standard curves and blanks).
4. Prepare working standards (S2–S8) by 1:3 (threefold) serial dilution.
Transfer the appropriate volume of standard into each of the labeled
tubes containing standard diluent as outlined above.
5. Vortex each standard at a medium setting for 5 sec before proceeding
with the next serial dilution. Change pipet tip at each dilution step.
Fig. 3. Preparing a threefold dilution series with a single reconstituted standard.
Reconstituted
Standard
100 100 100 100 100 100 100 100
S1 S2 S3 S4 S5 S6 S7 S8 Blank
Standard Diluent, µl
150 50 50 50 50 50 50 50 Transfer Volume, µl