Bio-Rad Bio-Plex Pro™ Human Th17 Cytokine Assays - Manual (Page 37)

35
Calculation Worksheet
If using either a premixed panel or one singleplex assay , follow these directions.
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 a 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 assay buffer required: _______ 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 a 20% excess to ensure enough volume: _______ µl x 0.2 = _______ µl
7 8
c. Total volume of 1x detection antibodies: _______ µl + _______ µl = _______ µl
7 8 9
d. Volume of 20x detection antibodies stock: _______ 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 10
b. Include 25% excess to ensure enough volume: _______ µl x 0.25 = _______ µl
10 11
c. Total volume of 100x streptavidin-PE: ______ µl + ______ µl = ______ µl
10 11 12
d. Volume of 100x streptavidin-PE required: _______ µl / 100 = _______ µl
12 13
e. Volume of assay buffer required: _______ µl – _______ µl = _______ µl
12 13 14