Bio-Rad Rat Diabetes Assays User Manual

Page 46

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If mixing diabetes assays (20x bead and detection antibody stocks) with cytokine assays
(10x stocks), follow these directions.
Note: Refer to Table 20 for the maximum number of diabetes and cytokine assays
that may be multiplexed. Mixing across panels is not applicable to NHP diabetes assays.

Enter the number of wells to be used in the assay:_______

1

Enter the number of diabetes tubes (either singleplex or multiplex) that will be mixed: ______

2

Enter the number of cytokine tubes (either singleplex or multiplex) that will be mixed: _______

3

1. Determine the volume of 1x diabetes and cytokine coupled beads needed.

a) Each well requires 50 µl of coupled beads (1x): _______ x 50 µl = _______ µl

1 4

b) Include 20% excess to ensure enough volume: _______ µl x 0.20 = _______ µl

4 5

c) Total volume of 1x coupled beads: _______ µl + _______ µl = _______ µl

4 5 6

d)

Volume of 20x beads required from each diabetes tube(s): _______ µl /20 = _______ µl

6 7

e) Volume of 10x beads required from each cytokines tube(s): ______ μl / 10 = ______ μl

6 8

f) Total volume of diabetes bead stock required: ______ μl x _______ = _______ μl

7 2 9

g) Total volume of cytokine bead stock required: ______ μl x _______ = _______ μl

8 3 10

h) Total volume of combined bead stocks required: _____ μl + ______ = _____ μl

9

10 11

i) Volume of assay buffer required: _______ µl - _______ µl = _______

6 11 12

Human, mouse, and rat diabetes

0

2

4

6

8

10

Mouse cytokine (group III) analytes (20x)

Human and mouse cytokine (groups I, II) analytes (10x)

10

9

8

7

6

5

Table 20. Maximum number of singleplex diabetes and cytokine analytes that may be multiplexed.

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