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.

Intracellular Staining

Related Protocols

Additional Tips

 

  • As cells can bind to antibodies non-specifically via their Fc receptors, using an Fc blocking reagent is highly recommended. For human samples, we recommend Human TruStain FcX™. For mouse samples, we suggest using TruStain FcX™ PLUS (anti-mouse CD16/32) antibody.
  • To prevent non-specific binding of cynanine-based dyes (i.e., PE/Cyanine7, APC/Cyanine7) to monocytes and macrophages, we recommend the use of True-Stain Monocyte Blocker™.
  • 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 Na2B4Owill 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.

BD Biosciences

BioLegend

559925 7-AAD Staining Solution 420403 7-AAD Viability Staining Solution
556463 Propidium Iodide 421301 Propidium Iodide
554724 BD™ GolgiStop 420701 Monensin Solution (1,000X)
555029 BD™ GolgiPlug 420601 Brefeldin A
556454 Annexin V Binding Buffer 422201 Annexin V Binding Buffer
555899 Lysing Buffer 420301 RBC Lysis Buffer (10X)
558049 Lyse/Fix Buffer 5X 422401 RBC Lysis/Fixation Solution (10X)
553141 Mouse Fc Block 156603 TruStain FcX™ PLUS (Mouse)
564219 BD Fc Block™ 422301 TruStain FcX™ (Human)
554656 Stain Buffer (FBS) 420201 Cell Staining Buffer
554722 BD Cytofix/Cytoperm™ Solution 420801 Fixation Buffer
554723 Perm/Wash Buffer 421002 Intracellular Staining Permeabilization Wash Buffer (10X)
560409/560098 (Mouse/Human) FOXP3 Buffer Set 424401 True-Nuclear™ Transcription Factor Buffer Set
338036 BD™ Stabilizing Fixative 422101 FluoroFix™ Buffer
None 422501 Cyto-Last™ Buffer
562574 Transcription Factor Buffer Set 424401 True-Nuclear™ Transcription Factor Buffer Set
558050 BD Phosflow™ Perm Buffer III 425401 True-Phos™ Perm Buffer
None 426101 True-Stain Monocyte Blocker™
554714 Fixation/Permeabilization Solution Kit 426803 Cyto-Fast™ Fix/Perm Buffer Set
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