Bio-Rad Human MMP and TIMP Assays User Manual

Page 26

Instrument

RP1 (PMT)

DD Gates

Bead Events

Bio-Plex 100, 200*

Low

5,000 (low), 25,000 (high)

Bio-Plex 3D*

Standard

Select MagPlex beads

Bio-Plex

MAGPIX

™

N/A, use default instrument settings

* A similar Luminex-based system may be used.

5. Click Enter Controls Info.
a. For user-specified controls, select an analyte from the dropdown

menu, then enter a description and concentration. Repeat for
each additional analyte in the assay.

For the vial of controls supplied, format the appropriate wells
as controls and enter descriptions but leave the concentrations
blank. Alternatively, the controls can be formatted as samples
with clear descriptions such as “control.” In any case, the
expected control ranges provided on the product data
sheet are not entered into Bio-Plex Manager software
version 6.1 and earlier.

6. Click Enter Sample Info and enter sample information and the

appropriate dilution factor.

7. Click Run Protocol and confirm that the settings follow Table 11.

Table 11. Read the plate using the appropriate instrument settings.

a. Confirm that data acquisition is set to 50 beads per region.

b. In Bio-Plex Manager software prior to 6.1, go to Advanced

Settings and confirm that the bead map is set to 100 region, the
sample size is set to 50 µl, and the doublet discriminator (DD) gates
are set to 5,000 (Low) and 25,000 (High). In Bio-Plex Manager
software versions 4.0, 4.1, 4.1.1, and 5.0, check Override Gates
and set the DD gate values as indicated.

Select Start, name and save the .rbx file, and begin data
acquisition. The Run Protocol pop-up screen will appear. Click
Eject/Retract to eject the plate carrier.