Bio-Rad Rat Diabetes Assays - Manual (Page 23)

21
# of Wells
96
20x
Beads, µl
288
Assay
Buffer, µl
5,472
Total
Volume, µl
5,760
# of Wells
96
20x beads, µl
Singleplex #1
288
20x beads, µl
Singleplex #2
288
Assay
Buffer, µl
5,184
Total
Volume, µl
5,760
# of Wells
96
20x
Beads, µl
Diabetes
288
10x
Beads, µl
Cytokines
576
Assay
Buffer, µl
4,896
Total
Volume, µl
5,760
5. Protect the beads from light with aluminum foil. Equilibrate to room
temperature prior to use.
Table 10 summarizes volumes required for preparing 1x beads from a
single 20x stock. Table 11 summarizes volumes required for preparing
1x beads from a mix of two 20x stocks; volumes required for preparing
beads of one diabetes (20x) assay and one cytokine (10x) assay (for
example, human insulin and human IL-6) are listed in Table 12.
Note: To minimize volume loss, use a 200–300 μl capacity pipet to
remove beads from the stock tube. If necessary, perform the volume
transfer in two steps. Do not use a 1,000 μl capacity pipet and/or
wide bore pipet tip.
Preparing 1x coupled beads from 20x stock (includes 20% excess volume).
Table 10. Premixed panel or one singleplex assay.
Table 11. Mixing two singleplex assays or a premixed panel + singleplex assay.
Table 12. Preparing 1x beads from two stocks at different concentrations. Mixing human
insulin (20x) with human IL-6 (10x) is one example*.
* Due to differences in dilution factors, it is not possible to multiplex adiponectin, adipsin
(human), VCAM-1, or ICAM-1 with other diabetes or cytokine assays.