Thermo Fisher/ebioscience
eBioscience Foxp3 / Transcription Factor Staining Buffer Set
00-5523-00
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We conduct flow cytometry experiments to analyze the differentiation process into subsets based on transcription factor (TF) analysis. In order to look at TFs in macrophages, we use this kit since it is a quick and easy way to perform both fixing and perming in one step before intracellular staining. The kit is easy to use and consistent.
Flow Cytometry
Murine Macrophages
Obtain single-cell suspension then proceed to surface stain your cells for 20–30 minutes at 4°C. Wash away the stain with FACS solution, and then mix 1 part of the concentrate to 3 part of the diluent (Fix/Perm) based on the number of samples. We typically use 100 ul of Fix/Perm for roughly max 5e6 total cells (scale up if needed). Fix/perm overnight at 4 deg or 30 minutes at room temperature away from light. Using 1x wash buffer, wash away the Fix/Perm before proceeding to intracellular protein staining.
Determine fixation/perm length conditions for optimal ICS staining. Too long can damage surface staining.
We are able to get robust transcription factor staining or cell cycle protein expression (Ki-67) using this kit.
N/A
None
Easy to use and very consistent
Great kit to fix and perm cells at once for intracellular protein staining of transcription factors or even cytokines