Bio-Rad Bio-Plex Pro™ Rat Cytokine, Chemokine, and Growth Factor Assays - Manual (Page 2)

6. Prepare a fourfold standard dilution series and blank as shown below.
Vortex for 5 sec between liquid transfers. If mixing diabetes assays with
cytokine assays, refer to the diabetes instruction manual.
7. After thawing samples, prepare as shown below.
8. Vortex the 10x or 20x coupled beads for 30 sec and dilute to 1x in
Bio-Plex assay buffer as shown. Protect from light.
Bio-Plex Pro Assay Quick Guide 4
Mouse Cytokine Group III and Rat Cytokine Group I Assays.
# of Wells
20x Beads, µl Assay Buffer, µl Total Volume, µl
96 288 5,472 5,760
Human and Mouse Cytokine Group I and II Assays.
# of Wells
10x Beads, µl Assay Buffer, µl Total Volume, µl
96 575 5,175 5,750
* If samples are serum-free, add BSA to 0.5% final w/v.
Serum and Plasma
Culture Supernatant
and Other Fluids
Cell and Tissue
Lysates
Dilution Dilution Dilution Diluent Diluent Diluent
Human, mouse,
and rat cytokines
Human
ICAM-1/VCAM-1
Mouse ICAM-1
1:4
1:100
1:200
User
optimized
User
optimized
User
optimized
User
optimized
(1:2 of
lysates at
200–900
µg/ml
protein)
Bio-Plex
sample
diluent
Bio-Plex
sample,
standard
diluent
Bio-Plex
serum-based
diluent
Cell
culture
medium
or buffer
similar to
sample*
Bio-Plex
sample
diluent
Assay
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
Note: Change tips between each dilution.