Bio-Rad Bio-Plex Pro™ Human Chemokine Assays - Manual (Page 15)

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4. During the incubation period, prepare the samples as instructed in
the Prepare Samples section.
Prepare the Standard Dilution Series
The following procedure produces an eight-point standard curve with a
fourfold dilution between each point. Pipet carefully using calibrated pipets
and use a new pipet tip for every volume transfer.
1. Label eight 1.5 ml polypropylene tubes S2 through S8 and Blank.
Alternatively, using Titertube ® micro test tubes may prove to be more
convenient if a multichannel pipet will be used to load the plate.
2. Add 150 µl of the appropriate diluent to tubes S2–S8 (Figure 3).
3. Vortex reconstituted standards at medium speed for 5 sec before
removing any volume. Transfer 50 µl to the S2 tube containing the
chosen standard diluent. Vortex for 5 sec.
4. Use a new pipet tip to transfer 50 µl from the S2 tube to the S3 tube.
Vortex for 5 sec.
5. Continue with 1:4 (fourfold) serial dilutions as shown in Figure 3.
6. Use reconstituted and diluted standards and controls immediately.
Do not freeze for future use.
Fig. 3. Preparing a fourfold dilution series with a single reconstituted standard.
150 150 150 150 150 150 150 150
S1 S2 S3 S4 S5 S6 S7 S8 Blank
Diluent, µl
50 50 50 50 50 50 50 Transfer Volume, µl
Reconstituted
Standard