Flow Cytometry Intracellular Fixation Permeabilization Buffer is designed for intracellular staining and flow cytometry analysis and is specially formulated for It is used to reduce non-specific staining of fluorescent dye-labeled antibodies and increase the fluorescence signal-to-noise ratio. In the first step, live cells can be "fixed" with a fixation buffer, which cross-links proteins. In the second step, membrane-breaking buffer is used to create pores in the membrane so that intracellular staining antibodies can effectively enter the cells. Permeabilization buffer should be used for subsequent washing steps, antibody addition, and incubation after cell rupture. Use Flow Cytometry Staining Buffer (Cat. No. 00-4222) to complete the final wash step and resuspend cells before running samples. This kit is commonly used to stain cytoplasmic proteins as well as proteins in the secretory pathway such as chemokines and cytokines. It is not recommended for intranuclear staining. For nuclear staining, see Foxp3/Transcription Factor Buffer Kit (Cat. No. 00-5523).

Before the fixation step, it is recommended to label viable cells with a Fixable Viability Dye (eFluor™ 450, 506, 660, 780, 520 or 455 UV) and then fix them.

Reported Applications
Flow cytometric analysis, intracellular staining followed by flow cytometric analysis