Bio-Rad Bio-Plex Pro™ Rat Cytokine, Chemokine, and Growth Factor Assays - Manual (Page 21)

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For the mouse ICAM-1 assay, dilute serum or plasma 1:200 in Bio-Plex
serum-based diluent, which is included in the Bio-Plex Pro high dilution
reagent kit, catalog #171-304080M.
For example:
n
First dilution (1:10)—10 µl sample + 90 µl diluent
n
Second dilution (1:20)—10 µl from the first dilution + 190 µl diluent
6. Prepare Coupled Beads
Instructions are provided for diluting the coupled beads to a 1x
concentration. When mixing cytokine and diabetes assays, keep in mind
the stock concentrations of coupled beads as listed in Table 9.
Note: When using 10-pack reagents, ensure that only the required volumes
of coupled beads, detection antibodies, streptavidin-PE, and buffers have
been removed from the tubes or bottles. For example, transfer a one-time
volume of assay buffer, sufficient to perform all steps of the assay procedure
(that is, prewetting the filter plate, diluting coupled beads, diluting
streptavidin-PE, and resuspending the beads) into a 50 ml reservoir.
1. Use Tables 10–14 to calculate the volume of coupled beads and
assay buffer needed.
2. Add the required volume of Bio-Plex assay buffer to a 15 ml
polypropylene tube.
Assay Stock Concentration of Coupled Beads
Human and mouse cytokines (groups I, II) 10x
Rat cytokines (group I) 20x
Mouse cytokines (group III) 20x
Human, mouse, and rat diabetes 20x
Table 9. Stock concentration of coupled beads.