Bio-Rad Bio-Plex Pro Human Acute Phase Reagent and Diluent Kits - Manual (Page 15)

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4. Gently vortex 1-3 sec and incubate on ice for 60 min. For optimal
assay performance, be consistent with the incubation time.
5. Save the standard/control diluents for preparation of the controls
(Section 8).
Prepare Standard Dilution Series
The standard 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 Standard
Dilution Series diagram on the next page.
2. Pipet the appropriate volume of standard diluent into the tubes (see
diagram below).
3. Refer to the steps and diagrams below for preparation of the
standard dilution series for the 5-plex and 4-plex assays.
5-plex standard dilution series:
Add 200 µl of reconstituted 5-plex
standard to the first 1.5 ml tube containing 200 µl 5-plex standard
diluent. Vortex gently. This mixture is identified as S1 in the diagram
below and in the standard and control values product insert
provided with the assay. Continue making serial dilutions of the
standards as shown in the diagram. Before making each dilution,
vortex gently and change the pipet tip.
5-plex Standard Dilution Series