The Mouse Inflammation Standards have been tested with BD™ Cytometric Bead Array (CBA) Mouse Inflammation Kit (Cat. No. 552364) to assure function as a standard in the CBA assay. Investigators are encouraged to refer to the manual for the BD™ Cytometric Bead Array (CBA) Mouse Inflammation Kit (Cat.No. 552364). The Mouse Inflammation Standards are lyophilized and should be reconstituted and serially diluted before mixing with CBA capture beads and PE detection reagents. An abbreviated instruction has been provided below (Preparation of Mouse Inflammation Standards). The Mouse Inflammation Standards should be reconstituted using Assay Diluent, which is provided with the BD™ Cytometric Bead Array (CBA) Mouse Inflammation Kit (Cat.No. 552364), or it may be purchased separately (Cat. No. 560104).
Preparation of Mouse Inflammation Standards for use with the BD™ CBA Mouse Inflammation Kit (Cat. No. 552364)
1. Open one vial of the lyophilized Mouse Inflammation Standards. Transfer all the lyospheres to a polypropylene tube (BD Falcon™, Cat. No. 352097). Label the tube "Top Standard".
2. Reconstitute the standard with 2.0 mL of Assay Diluent. Allow the reconstituted standard to equilibrate for at least 15 minutes before making dilutions. Mix the reconstituted protein by pipette only. Do not vortex or mix vigorously.
3. Label additional 12 x 75 mm tubes (BD Falcon™, Cat. No. 352008) and arrange them in the following order: Top Standard, 1:2, 1:4, 1:8, 1:16, 1:32, 1:64, 1:128, and 1:256.
4. Add 300 µl of Assay Diluent to each of the dilution tubes.
5. Perform a serial dilution by transferring 300 µl from the Top Standard to the 1:2 dilution tube and mix thoroughly. Do not vortex, mix by pipet only. 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. Prepare one tube containing only Assay Diluent to serve as the 0 pg/mL negative control.