Bio-Rad Bio-Plex Pro Human Acute Phase Reagent and Diluent Kits - Manual (Page 13)

9
5-plex and 4-plex assays
For the 5-plex assay, dilute 5 µl of
sample in 495 µl of serum-free diluent. Mix by vortexing gently
or pipetting. This results in a final 1:100 sample dilution. For the
4-plex assay, dilute 5 µl of the 1:100 mixture into an additional
495 µl of serum-free diluent. Mix gently. This results in a final
sample dilution of 1:10,000.
NOTE: Additional dilutions may be required in order for
samples with higher target concentrations to be within
assay range.
Culture Supernatant Samples
1. Collect and process the culture supernatant samples and assay
immediately or freeze at –20ºC. Avoid repeated freeze-thaw cycles.
2. If required, dilute the culture supernatant with culture medium.
Serum-free culture medium should contain carrier protein (such as
BSA) at a concentration of at least 0.5%. Keep the samples on ice until
ready for use.
Panel Step Dilution Sample Volume
Serum-free
Diluent Volume
Total
Volume
5-plex 1 100-fold 5 μl sample 495 μl 500μl
1 100-fold 5 μl sample 495 μl 500μl
4-plex
2 10,000-fold 5 μl of 1:100 495 μl 500μl