Bio-Rad Bio-Plex Pro™ TGF-β Assays - Manual (Page 15)

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Reconstitute a Single Vial of Standards
This procedure prepares enough material to run each dilution in duplicate.
1. Gently tap the vial containing the lyophilized standard.
2. Add 500 μl of the appropriate standard diluent (see Table 4). Do not
use assay buffer to reconstitute the standards.
3. Gently vortex the reconstituted standard for 5 sec, then incubate on
ice for 30 min. Be consistent with the incubation time in every assay
to ensure best results.
4. During the incubation period, prepare the samples as instructed in
section 5, Prepare Samples.
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 new pipet tips for every volume transfer.
1. Label nine 1.5 ml polypropylene tubes S1 through S8 and Blank.
2. Add the specified volume of standard diluent to each tube (Figure 3).
3. Vortex the reconstituted standards gently for 5 sec before removing
any volume. Add 128 µl to the S1 tube containing 72 µl of standard
diluent. Vortex at medium speed for 5 sec, then use a new pipet tip
to transfer 50 µl from S1 tube to S2 tube. Vortex.
4. Continue with 1:4 (fourfold) serial dilutions from tube S2 to S8 as
shown in Figure 3. Use reconstituted and diluted standards
immediately. Do not freeze for future use.
5. Continue with 1:4 serial dilutions as shown in Figure 3. Use
reconstituted and diluted standards immediately. Do not freeze for
future use.