Bio-Rad Bio-Plex Pro™ Rat Cytokine, Chemokine, and Growth Factor Assays - Manual (Page 17)

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4. Add 150 μl of standard diluent to the remaining tubes, as shown
in Figure 4.
Fig. 4. S1 mixture and fourfold dilution series of diabetes and cytokine standards for
detection at high PMT. Produces a narrow range cytokine standard curve.
5. For low PMT/broad range cytokine standard curve, add 64 µl of each
standard to the S1 tube, total volume 200 μl (Figure 5). Vortex for 5 sec.
6. Use a new pipet tip to transfer 50 μl from the S1 tube to the S2 tube
containing standard diluent. Vortex for 5 sec.
7. Continue with 1:4 (fourfold) serial dilutions from tube S2 to S8 as
shown in Figure 5, vortexing after each volume transfer.
Fig. 5. S1 mixture and fourfold dilution series of cytokine and diabetes standards for
detection at low PMT. Produces a broad range cytokine standard curve.
150 150 150 150 150 150 150 150
S2 S3 S4 S5 S6 S7 S8 Blank
Diluent (µl)
50 50 50 50 50 50 50
Transfer
Volume (µl)
250 µl
Reconstituted
Cytokine
Standard
72 µl
Standard Diluent
“S1” Tube, 200 µl Total
64 µl 64 µl
250 µl
Reconstituted
Diabetes
Standard
150 150 150 150 150 150 150 150
S2 S3 S4 S5 S6 S7 S8 Blank
Diluent (µl)
50 50 50 50 50 50 50
Transfer
Volume (µl)
500 µl
Reconstituted
Cytokine
Standard
59.2 µl
Standard Diluent
“S1” Tube, 200 µl Total
12.8 µl 128 µl
500 µl
Reconstituted
Diabetes
Standard