Bio-Rad Bio-Plex Pro Human Angiogenesis Reagent and Diluent Kits - Manual (Page 17)

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7. Prepare a sufficient volume of detection antibodies (1x) using
detection antibody diluent. Each well requires 2.5 µl of detection
antibodies (10x) adjusted to a final volume of 25 µl with detection
antibody diluent (refer to the example below).
8. After incubating the samples, slowly remove and discard the sealing
tape, then vacuum-filter.
9. Wash 3 times.
10. Vortex the detection antibodies gently and add 25 µl to each well.
Incubate for 30 min.
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 on the
following page).
Example Detection Antibody Calculations
# of Wells
10x Detection
Antibody (µl)
Total Volume (µl)
96
48
32
24
300
150
100
75
2,700
1,350
900
675
3,000
1,500
1,000
750
Detection Antibody
Diluent (µl)