Bio-Rad Bio-Plex Pro™ Human Chemokine Assays - Manual (Page 14)

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Selecting a Diluent for Standards and Controls
Refer to Table 4 for recommended diluents based on different sample types.
In order to meet the lot-specific control ranges provided on the product data
sheet, both the standards and controls should be reconstituted in Bio-Plex
®
standard diluent HB. If reconstituting in a different diluent, users will need to
establish/validate their own control ranges or acceptance criteria.
Reconstitute Standards and Quality Controls
This procedure prepares enough standard to run each dilution in duplicate.
Note: The appearance of the lyophilized standards or controls may vary
from a white pellet to clear crystals. Regardless of appearance, the vials
have passed QC specifications and perform accordingly.
1. Gently tap the vial containing the lyophilized standards on a solid
surface to ensure the pellet is at the bottom of the vial.
2. Reconstitute a single vial of standards with 781 µl of the appropriate
diluent. optional: at the same time, reconstitute the controls vial with
250 µl of the appropriate diluent as summarized in Table 4. Controls
do not require further dilution.
3. Vortex the reconstituted standards and controls at medium speed
for 5 sec, then incubate on ice for 30 min. It is important that
reconstitution of standards and controls is started and ended at the
same time. Be consistent with this incubation time to ensure optimal
assay performance and reproducibility.
Table 4. Summary of recommended diluents for standards and controls.
Diluent for Standards
Sample Type and Controls* Add BSA
Serum and plasma Standard diluent HB None
Culture media, with serum Culture medium None
Culture media, serum-free Culture medium To 0.5% final**
Lavage, sputum, other fluids Sample diluent HB To 0.5% final**
Lysate Sample diluent HB To 0.5% final**
* If using diluents other than standard diluent HB, users must establish their own control ranges.
** At least 0.5% final BSA is recommended to stabilize analytes and reduce adsorption to labware.