Bio-Rad Bio-Plex Pro™ TGF-β Assays - Manual (Page 2)

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.