Bio-Rad Bio-Plex Pro™ Human Isotyping 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 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 for 5 sec between liquid transfers.
Note: The quality controls are ready to use after reconstitution. No
dilution is needed.
8. After thawing samples, prepare according to the guidelines shown below.
9. Vortex coupled beads 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 Isotyping diluent None
Culture media, with serum Culture media None
Culture media, serum-free Culture media To 0.5% final
* If using diluents other than the isotyping diluent provided, then users must establish their
own control ranges.
Sample Type Diluent Add BSA Recommended Sample Dilution
Serum and plasma Isotyping None 1:40,000 for IgG
1
, IgG
2
, IgG
3
, IgG
4
, IgA, IgM
diluent 1:500 for IgE
1:20,000 for IgG total
Culture media, with serum Culture media None User defined
Culture media, serum-free Culture media To 0.5% final User defined
Bio-Plex Pro Assay Quick Guide 6