Bio-Rad Rat Diabetes Assays - Manual (Page 47)

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2. Determine the volume of 1x diabetes and cytokine detection antibodies needed.
a) Each well requires 25 μl of detection antibodies (1x): _______ x 25 μl = ______ μl
1 13
b) Include 25% excess to ensure enough volume: _______ μl x 0.25 = ______ μl
13 14
c) Total volume of 1x detection antibodies: ______ μl + ______ μl = ______ μl
13 14 15
d) Volume of 20x detection antibodies required from each diabetes tube(s):
______ μl / 20 = ______ μl
15 16
e) Volume of 10x detection antibodies required from each cytokines tube(s):
______ μl / 10 = ______ μl
15 17
f) Total volume of diabetes detection antibodies stock required: ____ μl x ____ =_____ μl
16 2 18
g) Total volume of cytokine detection antibodies stock required: ____ μl x ____ =_____ μl
17 3 19
h) Total volume of combined detection antibodies required:
_____ μl + ______ = _____ μl
18 19 20
i) Volume of detection antibody diluent required: ______ μl – _____ μl = _____ μl
15 20 21
3. Determine the volume of 1x streptavidin-PE needed.
a) Each well requires 50 μl of 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