The RayBio® Intracellular Staining Kit is used for assay of intracellular cytokine production in activated immune cells. Saponin-containing buffer commonly used to permeabilize cells and allow antibody to stain target intracellularly. With included Brefeldin A and Monensin, two inhibitors for Golgi apparatus function, cytokines will be trapped in cells and be detected by antibody in Cell Permeabilization Buffer.

• Fixation buffer (4% paraformaldehyde)
• Cell Permeabilization Buffer (10x)
• Brefeldin A (1000x)
• Monensin (1000x)

Kit can be stored at 4°C, protected from light for up to 3 months. Brefeldin A and Monensin can be stored as aliquots in -20°C for extended time.

One million mouse splenocytes were activated by Ionomycin (25 ng/ml) and PMA (1 µg/ml) for 5 hours in culture and stained with 0.25 µg RayBright® R647 anti-mouse CD4 and 0.5 µg RayBright® V450 anti-mouse IFNγ in 50 µl Cell Permeabilization Buffer.

Note: RayBright® is a trademark of RayBiotech, Inc.