Bio-Rad Human MMP and TIMP Assays - Manual (Page 14)

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Sample Type Diluent for Standard and Controls* Add BSA
Serum and plasma Diluent HD None
Culture media, with serum Culture media None
Culture media, serum-free Culture media To 0.5% final
* If using diluents other than the diluent HD provided, then users must establish their own
control ranges.
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
®
diluent HD. If reconstituting in a different diluent, users will need to establish/
validate their own control ranges or acceptance criteria.
Table 4. Summary of recommended diluents for samples.
Reconstitute Standards and 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 of lyophilized standards on a solid surface to
ensure that all material is at the bottom of the vial.
2. Reconstitute the vial of standards with 781 µl of the appropriate diluent.
optional: at the same time, reconstitute the vial of controls with 250 µl
of the appropriate diluent as summarized in Table 4. Controls do not
require further dilution. If using diluents other than the diluent HD
provided, then users must establish their own control ranges.
3. Gently vortex the reconstituted standards and controls 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.
4. During the incubation period, prepare the samples as instructed in the
Prepare Samples section.