Intracellular staining buffer selection

When performing intracellular staining for flow cytometry the selection of buffers used for fixation and permeabilization has a significant impact on the quality and accuracy of data. We provide optimized solutions for staining cytosolic, nuclear and secreted proteins for flow cytometry.

The Foxp3 Staining Buffer Set is optimized for staining all nuclear factors tested to date, including all transcription factors, whereas our conventional intracellular staining buffer system, Invitrogen eBioscience Fixation & Permeabilization is recommended for staining cytosolic and secreted proteins for flow cytometry.

We have investigated the compatibility of the Foxp3 Staining Buffer Set (cat. no. 00-5523) versus the Intracellular Fixation & Permeabilization Buffer Set (cat. no. 88-8824) for situations where simultaneous staining of both nuclear factors and cytosolic/secreted proteins is required.

Intracellular staining buffer selection

AntigenFoxp3 Staining Buffer SetFixation & permeabilization
HUMAN  
 IFNy * YES
 IL-1a nt YES
 IL-1b nt YES
 IL-2 * YES
 IL-4  * YES
 IL-5  nt YES
 IL-6 nt YES
 IL-9 nt YES
 IL-10  * YES
 IL-12 nt YES
 IL-13 nt YES
 IL-17A YES YES
 IL-17F YES YES
 IL-21 * YES
 IL-22 * YES
 Granzyme B nt YES
 Perforin nt YES
 TNFalpha YES YES
 Eomes YES NO
 Foxp3 YES NO
 Gata-3 YES NO
 RORy(t) YES NO
 T-Bet YES NO
 MOUSE  
 IFNy YES YES
 IL-1b nt YES
 IL-2 NO YES
 IL-4 * YES
 IL-6 nt YES
 IL-10 * YES
 IL-17A YES YES
 IL-17F YES YES
 IL-21 NO YES
 IL-22 * YES
 Granzyme B YES YES
 Perforin YES YES
 TNFalpha YES YES
 Eomes YES NO
 Foxp3 YES NO
 Gata-3 YES NO
 Nanog YES NO
 Oct 3/4 YES NO
 RORy(t) YES NO
 T-Bet YES NO

nt = Not tested

YES = Staining compatible with this buffer system

NO = Staining not compatible with this buffer system

*Staining is possible with the Foxp3 Staining Buffer Set, although in some cases a reduction in brightness has been observed. No change in the percentage of positive cells has been observed.

For Research Use Only. Not for use in diagnostic procedures.