Description
The RayBio? Fixation/Permeabilization Buffer contains paraformaldehyde, saponin and FBS, commonly used to fix and permeabilize cells simultaneously and allow further antibody staining intracellularly.
Buffer
PBS, PH7.2~7.4, 0.2% BSA, 0.1% NaN3
Cell Staining Protocol
- Activate cells in culture in the presence of Brefeldin A (1:1000 dilution in media) or monensin (1:1000 dilution in media).
- Wash the cells with PBS, spin down the cells at 500g, and then stain cells with live/dead markers and antibodies for surface markers.
- After surface staining, wash cells with cold PBS and incubate with RayBio Fixation/Permeabilization Buffer for 15-30 minutes at room temperature.
- Spin down the cells at 500g, wash the cells with 200 μl of Cell Permeabilization Buffer.
- Repeat step 4 once.
- Spin down the cells at 500g, resuspend cells in 50 μl of Cell Permeabilization Buffer containing antibody (antibodies) against cytokines or other intracellular proteins. Incubate for 30 minutes.
- Wash cells with 250 μl of Cell Permeabilization Buffer twice and resuspend cells in FACS buffer or 1% PFA in PBS.
- Analyze by flow cytometry.
Storage/Stability
Stored at 2 °C - 8 °C, protected from prolonged light exposure. DO NOT FREEZE.
Note
Note: RayBright? is a trademark of RayBiotech, Inc.
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