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

The Human Th1/Th2/Th17 Cytokine Standards have been tested with the BD™ Cytometric Bead Array (CBA) Human Th1/Th2/Th17 Cytokine Kit (Cat. No. 560484) to assure function as standards in the CBA assay. Investigators are encouraged to refer to the CBA kit manual or to the abbreviated instruction below (Preparation of Human Th1/Th2/Th17 Cytokine Standards Protocol). The Human Th1/Th2/Th17 Cytokine Standards should be reconstituted using Assay Diluent supplied with the aforementioned CBA kit (component no. 51-2432KC) or the standards may be purchased separately (Cat. No. 560104).

Preparation of Human Th1/Th2/Th17 Cytokine Standards: The Human Th1/Th2/Th17 Cytokine Standards are 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 Human Th1/Th2/Th17 Cytokine Standards. Transfer all the lyospheres to a 15 mL polypropylene tube (BD Falcon™, Cat. No. 352097). Label the tube "Top Standard".

2. Reconstitute the standards with 2.0 mL of Assay Diluent. Allow the reconstituted standards to equilibrate for at least 15 minutes before making dilutions. Mix the reconstituted proteins by pipet 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. ProClin is a trademark of Rohm and Haas Company.
2. This product is manufactured and sold under license from Pestka Biomedical Laboratories, Inc. (d/b/a PBL InterferonSource) and may be used solely as indicated. This product may not be resold or incorporated in any manner into another product for resale. Any use for therapeutics is strictly prohibited. This product is covered by U.S. Patent No. 5,597,901 and Bulgarian Patent No. BG1895.
3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
4. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
5. 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.