Bio-Rad Bio-Plex Pro Human Acute Phase Reagent and Diluent Kits - Manual (Page 22)

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11. While the detection antibodies are incubating, perform a 30 sec
quick-spin centrifugation of the streptavidin-PE (100x) prior to
pipetting to collect the entire volume at the bottom of the vial.
12. Prepare a sufficient volume of streptavidin-PE (1x) using assay buffer.
Each well requires 0.5 µl of streptavidin-PE (100x) adjusted to a final
volume of 50 µl with assay buffer (refer to the example below).
13. After the detection antibody incubation, slowly remove and discard
the sealing tape, then vacuum-filter.
14. Wash 3 times.
15. Vortex the streptavidin-PE (1x) vigorously and add 50 µl to each well.
Incubate for 10 min.
16. After the streptavidin-PE incubation, slowly remove and discard the
sealing tape, then vacuum-filter.
17. Wash 3 times.
18. Add 125 µl of assay buffer to each well. Cover the filter plate with a
new sheet of sealing tape. Shake the filter plate at room temperature
at 900 rpm for 30 sec and remove the sealing tape before
reading the plate.
# of Wells
100x
Streptavidin-PE
(µl)
Total Volume (µl)
96
48
32
24
60
30
20
15
5,940
2,970
1,980
1,485
6,000
3,000
2,000
1,500
Assay Buffer (µl)
Example Streptavidin-PE Calculations