Bio-plex pro assay quick guide 5 – Bio-Rad Bio-Plex Pro™ TGF-β Assays User Manual
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6. Reconstitute a single vial of standards in 500 µl of a diluent similar to the
final sample type or matrix as shown below. Vortex for 5 sec and incubate
on ice for 30 min.
7. Prepare a fourfold standard dilution series and blank as shown below.
Vortex for 5 sec between liquid transfers.
8. After thawing samples, activate by adding 1 volume of 1 N HCI to
5 volumes of sample. Vortex, incubate at RT for 10 min. Neutralize by
adding the same volume of base (1.2 N NaOH/0.5 M HEPES). After
treatment, dilute samples as shown below.
9. Vortex the 20x 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
Bio-Plex Pro Assay Quick Guide 5
Reconstituted
Standard
72 150 150 150 150 150 150 150 150
S1 S2 S3 S4 S5 S6 S7 S8 Blank
Diluent, µl
128 50 50 50 50 50 50 50
Transfer Volume, µl
Sample Type
Diluent for Standard
Add BSA
Serum and plasma
Standard/sample diluent mix (1:3)
None
Culture media, with serum
Culture media
None
Culture media, serum-free
Culture media
To 0.5% final (w/v)
Lavage, lysate, other fluids
Sample diluent
To 0.5% final (w/v)
Sample Type
Diluent
Add BSA
Sample Dilution
Serum and plasma
Sample diluent
None
1:16 final*
Culture media, with serum
Culture media
None
User optimized
Culture media, serum-free
Culture media
To 0.5% final
User optimized
Lavage, lysate, other fluids
Sample diluent
To 0.5% final
User optimized
* For example, activate 25 μl sample, neutralize, and bring to a final volume of 400 μl.