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

5. Reconstitute a single vial of standard in 781 µl of a diluent similar to your
final sample type or matrix. Reconstitute the quality controls in 250 µl of
the same diluent, as shown below. Vortex for 5 sec and incubate all vials
at once on ice for 30 min. (Section 4)
Note: The quality controls are ready to use after reconstitution. No dilution is
needed. Only premixed panels and x-Plex kits include controls.
6. Prepare a fourfold standard dilution series and blank as shown below.
Vortex for 5 sec between liquid transfers. (Section 4)
7. After thawing samples, prepare as shown below. (Section 5)
8. Vortex coupled beads for 30 sec and dilute to 1x in Bio-Plex assay buffer
as shown below. Protect from light. (Section 6)
# 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 Standard 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 standard diluent HB, users must establish their own control ranges.
Sample Type Diluent Add BSA 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, lysate, other fluids Sample diluent HB To 0.5% final User optimized
Bio-Plex Pro Assay Quick Guide 1