Thermo Fisher Scientific
Foxp3 / Transcription Factor Staining Buffer Set Kit
00-5523-00
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This kit is advertised for staining FoxP3 in T regulatory cells but it works well for all transcription factors and some cytokines. We use it to stain for FoxP3 in T regulatory cells but also Bcl-6 in T follicular cells and for perforin staining post stimulation. It can need some optimizing when working with a new stain but otherwise the product insert is a reliable protocol.
Flow Cytometry
Murine Lymphocytes and Splenocytes
Using a 1:4 dilution, mix fixation/permeabilization concentrate with fixation/permeabilization diluent. Add 250ul of fixation solution onto cells being fixed. Incubate at RT for 45 min or at 4 degrees overnight. Wash fixation solution away with 1X permeabilization buffer. Make 1x perm buffer from 10x stock solution diluted into water. Prior to staining I incubate cells for 10 min in 1x perm buffer and then apply desired stain for 45 min at room temperature.
You can use far less volume than what the protocol suggests for non-clinical samples.
You will get clean staining of any transcription factors including FoxP3 and Bcl-6. Optimization may be required for surface stains applied prior to fixation as this is a more intense fixation.
N/A
The best fixation buffer for FoxP3 staining
It can be too harsh for some surface stains so needs some optimization.
This fixation buffer works as advertised but may need some minor optimization for surface stains.