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Intracellular Staining Buffer Selection Guide

Intracellular Staining Buffer Selection

When performing intracellular staining for flow cytometry the selection of buffers used for fixation & permeabilization has a significant impact on the quality and accuracy of data. eBioscience provides optimized solutions for staining cytsolic, 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, 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  nt  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  YES  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.