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Preparation of Mouse Inflammation Standards
The Mouse Inflammation Standards are lyophilized and should be reconstituted
and serially diluted before mixing with the Capture Beads and the PE Detection
Reagent.
1. Open one vial of lyophilized Mouse Inflammation Standards. Transfer the
standard spheres to a polypropylene tube (eg, 15 ml Conical Tube,
BD Falcon™ Cat. No. 352097). Label tube "Top Standard".
2. Reconstitute the standards with 2.0 ml of Assay Diluent. Allow the
reconstituted standard to equilibrate for at least 15 minutes before making
dilutions.
Mix reconstituted protein by pipette only. Do not vortex or mix
vigorously.
3. Label 12 × 75 mm tubes (BD Falcon Cat. No. 352008) and arrange them
in the following order: 1:2, 1:4, 1:8, 1:16, 1:32, 1:64, 1:128, and 1:256.
4. Pipette 300 µl of Assay Diluent to each of the remaining tubes.
5. Perform a serial dilution by transferring 300 µl from the Top Standard to
the 1:2 dilution tube and mix thoroughly. Continue making serial dilutions
by transferring 300 µl from the 1:2 tube to the 1:4 tube and so on to the
1:256 tube and mix thoroughly (see Figure 2). Mix by pipette only, do not
vortex. Prepare one tube containing Assay Diluent to serve as the 0 pg/ml
negative control.
300 µ l 300 µ l 300 µ l 300 µ l 300 µ l 300 µ l 300 µ l 300 µ l
Top
Standard
1:2
2.0 mL
Dilution
T ube
1:4
Dilution
T ube
1:8
Dilution
T ube
1:16
Dilution
T ube
1:32
Dilution
T ube
1:64
Dilution
T ube
1:128
Dilution
T ube
1:256
Dilution
T ube
Figure 2. Preparation of Mouse Inflammation Standard Dilutions
The approximate concentration (pg/ml) of recombinant protein in each dilution
tube is shown in Table 1.
