BioLegend Flow Cytometry Buffers
Beyond antibody reagents, flow cytometry requires the right types of buffers for optimal staining. This convenient list separates out flow cytometry applications by their intended target. It also indicates which buffers are best-suited to your task for surface or intracellular staining and the protocols necessary for each.
Surface & Intracellular Staining Equivalent Products Breaking Buffers
Surface Staining
 Tissue/Cells/Whole BloodApoptosis (Annexin V)
Surface StainingCell Staining BufferAnnexin Binding Buffer
FixationFixation BufferFixation Buffer
Viability Dyes

7-AAD Viability Staining Solution

Propidium Iodide

DRAQ7™

Helix NP™ Dyes

7-AAD Viability Staining Solution

Propidium Iodide

DRAQ7™

Helix NP™ Dyes

Additional Tips
  • You may have to check if other surface markers are sensitive to the addition of the calcium present in the Annexin Binding Buffer.
Intracellular Staining
 CytokinesFOXP3/Helios/
Transcription Factors
Phospho TargetsBrdU
Surface StainingCell Staining BufferCell Staining BufferCell Staining BufferCell Staining Buffer
Fixation

Fixation Buffer

Cyto-Fast™ Fix/Perm Buffer Set

True-Nuclear™ Transcription Factor Buffer SetFixation BufferPhase-Flow™ BrdU Kits
Permeabilization

Intracellular Staining Permeabilization Wash Buffer (10X)

Cyto-Fast™ Fix/Perm Buffer Set

True-Nuclear™ Transcription Factor Buffer SetTrue-Phos™ Permeabilization BufferPhase-Flow™ BrdU Kits
Viability DyesZombie DyesZombie DyesZombie Dyes

7-AAD Viability Staining Solution

DAPI

Additional Tips
  • For phospho staining, we recommend using RBC Lysis/Fixation Solution (10X) if the sample type is unlysed mouse spleen or whole blood. In addition, DAPI or DRAQ5™ may be used as a nucleic acid staining reagent, but only after the fixation/permeabilization step.
  • Zombie Dyes should be stained prior to any other antibody stain in the protocol as it will bind to any available primary amines.
  • For activation of cytokine production in cell culture, we recommend our Activation Cocktails, containing PMA and ionomycin, with or without Brefeldin A.
Red Blood Cell Lysis
RBC Lysis Buffer (10X)
RBC Lysis/Fixation Solution (10X)
Special Buffer Applications
BufferUtility
Human TruStain™ FcXBlocks unwanted Fc receptor binding of antibodies on human cells.
Mouse TruStain™ FcX PLUS

Blocks unwanted Fc receptor binding of antibodies on mouse cells. Shows improved blocking ability over the previous Mouse TruStain™ FcX antibody (clone 93).

True-Stain Monocyte Blocker™

Prevents monocytes and macrophages from binding non-specifically to fluorophores. This binding occurs independently of Fc blocking.

FluoroFix™ Buffer Provides a gentler fixation for flow cytometry samples; particularly useful for tandem dyes. 
Cyto-Last™ BufferAllows cells that cannot be analyzed immediately for cytokine staining to be stored for up to two weeks.
Flow Cytometry Antibody Diluent BufferRecommended for the preparation of concentrated antibodies or staining cocktails.
Related Protocols
RBC Lysis/Fixation
Cell Surface Immunofluorescence Staining Protocol
Intracellular Cytokine Staining
Intracellular Cytokine Stimulation Guide
True-Nuclear™ Transcription Factor Staining Protocol
Intracellular Staining with True-Phos™ Perm Buffer in Whole Blood
Intracellular Staining with True-Phos™ Perm Buffer in Cell Suspensions
Phase-Flow™ BrdU Kit Manual
Ethanol Fixation
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