BD™ Cytometric Bead Array (CBA) Mouse Th1/Th2/Th17 Cytokine Standards

The Mouse Th1/Th2/Th17 Cytokine Standard has been tested with BD™ Cytometric Bead Array (CBA) Mouse Th1/Th2/Th17 Cytokine Kit (Cat. No. 560485) to assure function as a standard in the CBA assay.  Investigators are encouraged to refer to the CBA kit manual or to the brief instructions below (Preparation of Mouse Th1/Th2/Th17 Cytokine Standard Protocol).  The Mouse Th1/Th2/Th17 Cytokine Standard should be reconstituted using Assay Diluent supplied with the aforementioned CBA kit (component no. 51-2432KC) or it may be purchased separately (Cat. No. 560104).

Preparation of Mouse Th1/Th2/Th17 Cytokine Standard:  The Mouse Th1/Th2/Th17 Cytokine Standard is lyophilized and should be reconstituted and serially diluted before mixing with CBA capture beads and PE detection reagents.

1. Open one vial of the lyophilized Mouse Th1/Th2/Th17 Cytokine Standard. Transfer all the lyospheres to a 15 mL 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.

1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
3. ProClin is a trademark of Rohm and Haas Company.
4. Warning: CBA lyophilized standard contains 0.02% (w/w) and Detection Reagent (Part A) contains 0.002% (w/w) of a CMIT/MIT mixture (3:1), which is a mixture of: 5-chloro-2-methyl-4-isothiazolin-3-one [EC No 247-500-7] and 2-methyl-4-isothiazolin-3-one [EC No 220-239-6] (3:1).Hazard statement: May cause an allergic skin reaction.Precautionary statements: Contaminated work clothing should not be allowed out of the workplace. Wear protective gloves/eye/face protection. Wear protective clothing. Avoid breathing mist/vapours/spray. If skin irritation or rash occurs: Get medical advice/attention. IF ON SKIN: Wash with plenty of water. Take off contaminated clothing and wash it before reuse. Dispose of contents/container in accordance with local/regional/national/international regulations.