Bio-Rad Bio-Plex Pro™ Human Th17 Cytokine Assays - Manual (Page 27)

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4. Enter Standards Info into Bio-Plex Manager.
a. Enter the highest concentration of each analyte in the top row
(labeled S1) of the table. S1 concentration information is included
with each vial of standards.
b. Enter a dilution factor of 4 and click Calculate. The concentrations
for each standard point will be populated for all analytes in the table.
c. Optional: enter the lot number of the vial of standards into the
Standard Lot box and click Save.
5. Enter Controls Info — for user specified controls, select an analyte
from the pull-down menu, then enter a description and concentration.
Repeat for each additional analyte in the assay.
For the quality controls supplied in premixed Th17 cytokine kits,
format the appropriate wells as controls, enter descriptions, but leave
the concentrations blank. Alternatively, the quality controls can be
formatted as samples with clear descriptions such as “quality control
high” and “quality control low”. In any case, the expected control
ranges provided on the product data sheet are not entered into
Bio-Plex Manager software v6.1 and earlier.
6. Enter Sample Info — enter sample information and the appropriate
dilution
factor.
7. Run Protocol — confirm that the assay settings are correct.
a. Human Th17 cytokine assays are run at the default setting in
Bio-Plex Manager — low RP1 target (low PMT).
b. Confirm that data acquisition is set to 50 beads per region.
In Advanced Settings, confirm that the bead map is set to 100
region, the sample size is set to 50 µl, and the 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.