Bio-Rad Bio-Plex Pro Human Acute Phase Reagent and Diluent Kits - Manual (Page 21)

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6. While the samples are incubating, perform a 30 sec quick-spin
centrifugation of the detection antibody (10x for multiplex assays,
100x for singleplex assays) prior to pipetting to collect the entire
required volume at the bottom of the vial.
7. Prepare a sufficient volume of detection antibodies (1x) using
detection antibody diluent. Refer to the example tables below.
NOTE: The detection antibody stock concentration is 10x in the
multiplex assays and 100x in the singleplex assays. Always check
the detection antibody concentration on the vial label before diluting.
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.
Example Detection Antibody Calculations (10x stock)
# 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)
Example Detection Antibody Calculations (100x stock)
# of Wells
100x Detection
Antibody (µl)
Total Volume (µl)
96
48
32
24
30
15
10
7.5
2,970
1,485
990
742.5
3,000
1,500
1,000
750
Detection Antibody
Diluent (µl)