Bio-Rad Bio-Plex® Precision Pro™ Human Cytokine Assays - Manual (Page 9)

Section 6
Standard Preparation
Two tubes of Iyophilized cytokine standard are provided in each Bio-Plex ®
Precision Pro™ cytokine assay. However, only one of the tubes is required per
96-well plate. The product insert provided with the assay lists the
concentration of the reconstituted standard. This procedure will prepare
enough standard to run each dilution in duplicate.
Reconstitute Standards
1. Gently tap the glass vial containing the lyophilized cytokine standard
on a solid surface to ensure the pellet is at the bottom.
2. Reconstitute 1 vial of lyophilized standard with 500 µl of the
appropriate standard diluent. Do not use assay buffer to dilute
standards.
3. Gently vortex 1–3 sec and incubate on ice for 30 min. Be
consistent with the incubation time for optimal assay performance.
Prepare Standard Dilution Series
The cytokine concentrations specified for the 8-point standard dilution set
have been selected for optimized curve fitting using the 5-parameter
logistic (5PL) or 4-parameter logistic (4PL) regression in Bio-Plex
Manager™ software. Results generated using dilution points other than
those listed in this manual have not been optimized.
1. Label a set of 1.5 ml Eppendorf tubes as shown in the diagram on
the next page.
Serum Serum standard diluent
Plasma Plasma standard diluent
Culture supernatant Same culture medium used
to prepare samples
Sample Standard Diluent
7