Intracellular staining is essential to identifying unique cell subsets and characterizing cell populations. Intracellular flow cytometry is commonly used to analyze intracellular molecules including cytokines, transcription factors, inflammatory mediators, and even phosphorylated proteins.

The RayBiotech Intracellular Staining Kit is designed to allow for intracellular staining of target cytokines, proteins, or other factors in activated or treated cell populations. The kit includes both Brefeldin A and Monensin both of which inhibit the Golgi apparatus function, trapping normally secreted cytokine and proteins inside cells during incubation. Following incubation, the included saponin-containing buffer is used to permeabilize cells in order to allow intracellular antibodies entry through the plasma membrane.

Intracellular staining must be done using a permeabilization buffer to allow antibodies to have access to the cytoplasm of the cell. Once allowed in, antibodies of choice can be used to target specific proteins and characterize the cell population. This provides the researcher with abundant information about cellular function and signaling mechanisms.

Product Analysis

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.