Bio-Rad Bio-Plex Pro™ Human Th17 Cytokine Assays - Manual (Page 14)

12
Reconstitute Standards and Quality Controls
This procedure prepares enough standard to run each dilution in duplicate.
1. Gently tap the vial containing the lyophilized standards on a solid
surface to ensure the pellet is at the bottom of the vial.
2. Reconstitute a single vial of standards with 781 µl of the appropriate
diluent. Optional: at the same time, reconstitue the two control vials
with 250 µl of the appropriate diluent as summarized in Table 4.
Controls do not require further dilution.
3. Gently vortex the reconstituted standards and controls for 5 sec,
then incubate on ice for 30 min. It is important that reconstitution
of standards and controls is started and ended at the same time.
Be consistent with this incubation time to ensure optimal assay
performance and reproducibility.
4. During the incubation period, prepare the samples as instructed in the
Prepare Samples section.
Prepare the Standard Dilution Series
The following procedure produces an eight-point standard curve with a
four-fold dilution between each point. Pipet carefully using calibrated pipets
and use new pipet tips for every volume transfer.
1. Label eight 1.5 ml polypropylene tubes S2 through S8 and Blank.
2. Add 150 µl of the appropriate diluent to tubes S2–S8 (Figure 3).
3. Vortex reconstituted standards gently for 5 sec before removing
any volume. Add 50 µl to the S2 tube containing the chosen standard
diluent. Vortex for 5 sec.