Bio-Rad Rat Diabetes Assays - Manual (Page 17)

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3. For high PMT/narrow range cytokine standard curve, add 12.8 μl
of the reconstituted cytokine standard and 128 μl of the
reconstituted diabetes standard to the S1 tube for a total volume
of 200 μl (Figure 4). Vortex at medium speed for 5 sec.
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.
7. Continue with 1:4 (fourfold) serial dilutions from tube S2 to S8 as
shown in Figure 5.
Fig. 5. S1 mixture and fourfold dilution series of diabetes and cytokine 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