Bio-Rad Bio-Plex Pro™ Human Chemokine Assays - Manual (Page 2)

6. Reconstitute the vial of standards in 781 µl of a diluent similar to your final
sample type or matrix. Reconstitute the vial of quality controls in 250 µl of
the same diluent, as shown below. Vortex at medium speed for 5 sec and
incubate all vials at once on ice for 30 min.
7. Prepare a fourfold standard dilution series and blank as shown below.
Vortex at medium speed for 5 sec between liquid transfers.
Note: The quality controls are ready to use after reconstitution. No
dilution is needed. Quality controls are included with the fixed panel only.
8. After thawing samples, prepare according to the guidelines shown below.
9. Vortex coupled beads at medium speed for 30 sec and dilute to 1x in
Bio-Plex assay buffer as shown below. Protect from light.
# of Wells 20x Beads, µl Assay Buffer, µl Total Volume, µl
96 288 5,472 5,760
150 150 150 150 150 150 150 150
S1 S2 S3 S4 S5 S6 S7 S8 Blank
Diluent, µl
50 50 50 50 50 50 50 Transfer Volume, µl
Reconstituted
Standard
Sample Type Diluent for Standards and Controls* Add BSA
Serum and plasma Standard diluent HB None
Culture media, with serum Culture media None
Culture media, serum-free Culture media To 0.5% final
Lavage, lysate, other fluids Sample diluent HB To 0.5% final
* If using diluents other than the standard diluent HB provided, then users must establish
their own control ranges.
Sample Type Diluent Add BSA Recommended Sample Dilution
Serum and plasma Sample diluent HB None Fourfold (1:4)
Culture media, with serum Culture media None Neat to 1:10
Culture media, serum-free Culture media To 0.5% final Neat to 1:10
Lavage, other fluids Sample diluent HB To 0.5% final User optimized
Lysate Sample diluent HB To 0.5% final User optimized (at least 1:2 for
50 to 500 μg/ml final protein)
Bio-Plex Pro Human Chemokine Quick Guide