Bio-Rad Bio-Plex Pro Human Acute Phase Reagent and Diluent Kits - Manual (Page 14)

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Section 7
Standard Preparation
Two vials of Iyophilized acute phase standards are provided in each
multiplex Bio-Plex Pro™ acute phase assay kit and one vial is provided in
each singleplex assay kit. Only one vial is required per 96-well plate. The
product insert provided with the assay lists the concentration of the most
concentrated dilution point of the standard curve. This procedure will
prepare enough standard to run each dilution in duplicate.
Reconstitute Standards
1. Reconstitute one 5-plex standard for every 5-plex assay and one 4-
plex standard for every 4-plex assay. Gently tap the glass vial
containing the lyophilized acute phase standards on a solid surface
to ensure that the pellet is on the bottom.
2. Refer to the following steps for preparation of diluents for the 5-plex
and 4-plex standards and controls. Use the serum-based diluent and
serum-free diluent provided in the Bio-Plex
® acute phase diluent kit
(catalog# 171-305050) for serum and plasma samples. For tissue
culture samples, use tissue culture medium. Do not use assay buffer
to dilute standards and controls.
5-plex standard/control diluent:
Dilute 1 ml of serum-based
diluent in 24 ml of serum-free diluent. Vortex 10 sec. Label this
mixture "5-plex standard/control diluent".
4-plex standard/control diluent:
The serum-free diluent is
used directly for diluting 4-plex standards and controls. Using a
sterile pipette, transfer 25 ml into a clean container labeled "4-
plex standard/control diluent".
3. Reconstitute 1 standard vial per assay with 0.5 ml of the appropriate
diluent as described in Step 2. The mixtures are identified as S0 in
the Standard Dilution Series diagrams on page 11.