Bio-Rad Bio-Plex Pro™ Rat Cytokine, Chemokine, and Growth Factor Assays - Manual (Page 18)

16
5. Prepare Samples
General guidelines for preparing different sample types are provided here.
For more information, contact Bio-Rad Technical Support.
n
Once thawed, keep samples on ice. Prepare dilutions just prior to the
start of the assay and equilibrate to room temperature before use
n
Do not freeze diluted samples
Table 8. Summary of recommended sample diluents and dilution factors.
Sample Preparation
Serum and Plasma
EDTA or citrate is preferred as an anticoagulant. Heparin-treated plasma,
while compatible with Bio-Plex Pro
™ assays, may absorb certain soluble
proteins of interest. Avoid using hemolyzed samples as this may lead to
false positive results.
1. Draw whole blood into collection tubes containing anticoagulant.
Invert tubes several times to mix.
Serum and Plasma
Culture Sup and
Other Fluids
Cell and Tissue
Lysates
Dilution Dilution Dilution Diluent Diluent Diluent
* Requires a two-step dilution scheme as described on pages 18–19. Due to differences in
dilution factors, it is not possible to multiplex ICAM-1 or VCAM-1 with other assays.
** If samples are serum-free, add BSA to at least 0.5% final w/v to stabilize analytes and reduce
adsorption to labware.
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
(at least
1:2 of
lysates at
200–900
ug/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