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
| Antigen | Foxp3 Staining Buffer Set | Fixation & 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.
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For Research Use Only. Not for use in diagnostic procedures.