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Calculation worksheet – Bio-Rad Bio-Plex Pro™ Magnetic Cell Signaling Assays User Manual

Page 34

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32

Calculation Worksheet

If using either a

premixed panel or one singleplex assay

, follow these instructions.

Plan the plate layout and 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 of 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. Volume of 20x coupled beads stock: _______ µl/20 = _______ µl

4 5

e. Volume of wash buffer required: _______ µl – _______ µl = _______ µl

4 5 6

2. Determine the volume of 1x detection antibody needed.
a. Each well requires 25 µl detection antibodies (1x): ______ x 25 µl = _______ µl

1 7

b. Include 25% excess to ensure enough volume: _______ µl x 0.25 = _______ µl

7 8

c. Total volume of 1x detection antibodies: _______ µl + _______ µl = _______ µl

7 8 9

d. Volume of 20x detection antibodies required: _______ µl/20 = _______ µl

9 10

e. Volume of detection antibody diluent required: _____ µl – _____ µl = _____ µl

9 10 11

3. Determine the volume of 1x streptavidin-PE needed.
a. Each well requires 50 µl streptavidin-PE (1x): _______ x 50 µl = _______ µl

1 12

b. Include 25% excess to ensure enough volume: _______ µl x 0.25 = _______ µl

12 13

c. Total volume of 100x streptavidin-PE: ______ µl + ______ µl = ______ µl

12 13 14

d. Volume of 100x streptavidin-PE required: _______ µl/100 = _______ µl

14 15

e. Volume of detection antibody diluent required: _______ µl – _______ µl = _______ µl

14 15 16