Bio-Rad Rat Diabetes Assays - Manual (Page 44)

42
Calculation Worksheet
If using either a premixed panel or one singleplex assay with 20x stocks of beads
and detection antibodies, 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 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 required: _______ 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 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 10
b. Include 25% excess to ensure enough volume: _______ µl x 0.25 = _______ µl
10 11
c. Total volume of 1x 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