Bio-Rad Bio-Plex Pro Human Acute Phase Reagent and Diluent Kits - Manual (Page 16)

4-plex standard dilution series:
Add 50 µl of reconstituted 4-plex
standard to the first 1.5 ml tube containing 350 µl 4-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. Keep the standards on ice until ready for use. Standards should be
used immediately and should not be frozen for future use.
12
4-plex Standard Dilution Series
Note: The addition of duplicate 0 pg/ml blanks is strongly
recommended. Use 50 µl of the appropriate standard diluent as the
blank sample. the 0 pg/ml points should be formatted as blanks, not
as points in the curve, when using Bio-Plex Manager software.
Formatting the 0 pg/ml wells as blanks results in an automatic
subtraction of the background Median Flourescence Intensity (MFI)
values from all readings in a run. This might result in negative MFI
values in some wells. If this is undesirable, the 0 pg/ml wells can be
formatted as background controls. The 0 pg/ml background control
wells are also useful for troubleshooting and determining the limit of
detection (LOD).