Flow cytometric analysis of mouse splenocytes fixed and permeabilized with #43481 FoxP3/Transcription Factor Fixation/Permeabilization kit, using FoxP3 (3G3) Mouse mAb (PE Conjugate) #49402 (right) and co-stained with CD4-APC #78292, compared to a concentration-matched Mouse (MOPC-21) mAb IgG1 Isotype Control (PE Conjugate) #63630 (left).
Store at 4ºC. All components in this kit are stable for at least 6 months when stored at 4ºC.
NOTE: Precipitation may occur in some kit components. The presence of precipitate does not affect the performance of the reagent.
Additional Reagents (Not Supplied):
NOTE: Fixable viability dyes such as Ghost Dye(TM) Violet 510 Viability Dye #59863 should be added prior to fixation, following the dye product recommended protocol. Proceed with fixation once excess dye has been removed.
NOTE: Antibodies targeting CD markers or other extracellular epitopes may be added prior to fixation. The antibodies will remain bound to the target of interest during the fixation process. A wash step prior to fixation may be performed but is not necessary.
NOTE: Optimal centrifugation conditions will vary depending upon cell type and reagent volume. Generally, 150-300g for 1-5 minutes will be sufficient to pellet the cells.
revised June 2020
Protocol Id: 1744
This kit provides the reagents needed to support antibody-based detection of FoxP3 and other transcription factors by flow cytometry. Optimally formulated fixation and permeabilization buffers halt biological activity and enable antibody access to intracellular targets. Antibody dilution, incubation, and wash steps are performed with the included FoxP3/Transcription Factor Permeabilization Buffer, eliminating the need for separate reagents.
Note: Precipitation may occur in some kit components. The presence of precipitate does not affect the performance of the reagent.
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