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 Staining420201 Cell Staining Buffer422201 Annexin Binding Buffer
Fixation420801 Fixation Buffer420801 Fixation Buffer
Viability Dyes420403 7-AAD Viability Staining Solution420403 7-AAD Viability Staining Solution
 421301 Propidium Iodide421301 Propidium Iodide
 424001 DRAQ7™424001 DRAQ7™
 425301 Helix NP™ NIR425301 Helix NP™ NIR
Intracellular Staining
 CytokinesFOXP3/Helios/
Transcription Factors
Phospho TargetsBrdU
Surface Staining420201 Cell Staining Buffer420201 Cell Staining Buffer420201 Cell Staining Buffer420201 Cell Staining Buffer
Fixation420801 Fixation Buffer424401 True-Nuclear™ Transcription Factor Buffer Set420801 Fixation Buffer420801 Fixation Buffer
Permeabilization421002 Intracellular Staining Permeabilization Wash Buffer (10X)424401 True-Nuclear™ Transcription Factor Buffer Set425401 True-Phos™ Permeabilization Buffer0.5% Triton-X 100 in PBS
Viability DyesZombie DyesZombie DyesZombie Dyes420403 7-AAD Viability Staining Solution
    422801 DAPI
Red Blood Cell Lysis
420301 RBC Lysis Buffer (10X)
422401 RBC Lysis/Fixation Solution (10X)
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
Anti-BrdU Staining Protocols
Ethanol Fixation
Additional Tips
  • Please note the BrdU buffer reagents listed above reflect the protocol utilizing DNAse. If the ethanol treatment is preferred, 70% ethanol, 2N HCl, and 0.1M Na2B4O7 will be required instead. See the BrdU staining protocol for more information.
  • You may have to check if other surface markers are sensitive to the addition of the calcium present in the Annexin Binding Buffer.
  • For gentler fixation, Cat# 422101 FluoroFix™ Buffer can be used.
  • Cat# 425501 Flow Cytometry Antibody Diluent Buffer is recommended for the preparation of concentrated antibodies or staining cocktails.
  • Cells that cannot be analyzed immediately for cytokine staining can be stored in Cat# 422501 Cyto-Last™ Buffer.
  • For phospho staining, we recommend using Cat# 422401 RBC Lysis/Fixation Solution (10X) if the sample type is unlysed mouse spleen or whole blood.
  • Zombie Dyes should be stained prior to any other antibody stain in the protocol as it will bind to any available primary amines.
  • For Phospho Targets, Cat# 422801 DAPI or Cat#424101 DRAQ5™ may be used as a nucleic acid staining reagent, but only after the fixation/permeabilization step.
  • To prevent non-specific binding of cynanine-based dyes (i.e., PE/Cy7, APC/Cy7) to monocytes and macrophages, we recommend the use of True-Stain Monocyte Blocker™.
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