Bio-Rad Bio-Plex Pro™ Rat Cytokine, Chemokine, and Growth Factor Assays User Manual
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If mixing singleplex assays with 20x stocks of beads and detection antibodies, follow
these directions. If using 10x stocks, divide by 10 instead of 20 in steps 1e and 2e.
Enter the number of wells to be used in the assay: _______
1
1. Determine the volume of 1x coupled beads needed.
a. Each well requires 50 µl coupled beads (1x): _______ x 50 µl = _______ µl
1 2
b. Include 20% excess to ensure enough volume: _______ µl x 0.20 = _______ µl
2 3
c. Total volume of 1x coupled beads: _______ µl + _______ µl = _______ µl
2 3 4
d. Enter the number of singleplex sets (or analytes) tubes that will be multiplexed = _______
5
e. Volume of 20x coupled beads required from each stock tube:
_______ µl / 20 = _______ µl
4 6
f. Total volume of combined bead stocks: _______ x _______ µl = _______ µl
5 6
7
g. Volume of assay buffer required: _______ µl – _______ µl = _______ µl
4 7 8
2. Determine the volume of 1x detection antibody needed.
a. Each well requires 25 µl detection antibodies (1x): _______ x 25 µl = _______ µl
1 9
b. Include 25% excess to ensure enough volume: _______ µl x 0.25 = _______ µl
9 10
c. Total volume of 1x detection antibodies: _______ µl + _______ µl = _______ µl
9
10 11
d. Enter the number of singleplex sets (or analytes) that will be multiplexed = _______
5
e. Volume of 20x detection antibodies required from each stock tube:
_______ µl / 20 = _______ µl
11 12
f. Total volume of combined detection antibody stock: _____ µl x _____ = _____ µl
12
5 13
g. Volume of detection antibody diluent required: ____ µl – ____ µl = ____µl
11 13 14
3. Determine the volume of 1x streptavidin-PE needed.
a. Each well requires 50 µl streptavidin-PE (1x): _______ x 50 µl = _______ µl
1 15
b. Include 25% excess to ensure enough volume: _______ µl x 0.25 = _______ µl
15 16
c. Total volume of 1x streptavidin-PE: ______ µl + ______ µl = _______ µl
15 16 17
d. Volume of 100x streptavidin-PE required: _______ µl / 100 = _______ µl
17 18
e. Volume of assay buffer required: _______ µl – _______ µl = _______ µl
17 18 19