Bio-Rad Bio-Plex Pro™ Rat Cytokine, Chemokine, and Growth Factor Assays - Manual (Page 29)

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8. Cover plate with a new sheet of sealing tape and protect from light
with aluminum foil. Incubate on shaker at 850 ± 50 rpm for 10 min at
room temperature.
9. After the streptavidin-PE incubation step, slowly remove and discard
the sealing tape.
10. Wash the plate three times with 100 µl wash buffer.
11. To resuspend beads for plate reading, add 125 µl of assay buffer to
each well. Cover the plate with a new sheet of sealing tape. Shake at
room temperature at 850 ± 50 rpm for 30 sec, and slowly remove the
sealing tape. Ensure that the plate cover has been removed before
placing the plate on the reader.
12. Remove the sealing tape and read the plate using the settings
below. Refer to section 8 for details.
Note: Reading at alternative PMT settings on the Bio-Plex 100,
200, or Bio-Plex 3D (for example when mixing diabetes assays with
cytokine assays) requires validation by the end user to ensure that
results meet the user’s acceptance criteria.
Table 24. Read the plate using the appropriate instrument settings.
RP1 (PMT) Setting
Bio-Plex Pro Assay Bio-Plex 100/200* Bio-Plex 3D* Bio-Plex ® MAGPIX ™ *
Human cytokines (group l, ll) Low
Mouse cytokines (group l, ll, III) Low Standard
Rat cytokines (group l) High
Cross-Panel Mixing
Human cytokines + diabetes
Mouse cytokines + diabetes High Standard
Rat cytokines + diabetes
Other Instrument Settings
DD gates 5,000–25,000 Select MagPlex
beads
Bead events 50 50
* Or similar Luminex-based system.
N/A,
Use
default
settings