Surface Staining

  Tissue/Cells/Whole Blood Apoptosis (Annexin V)
Surface Staining Cell Staining Buffer Annexin Binding Buffer
Fixation Fixation Buffer Fixation Buffer
Viability Dyes 7-AAD Viability Staining Solution 7-AAD Viability Staining Solution
  Propidium Iodide Propidium Iodide
  DRAQ7™ DRAQ7™
  Helix NP™ NIR Helix NP™ NIR

Intracellular Staining

Red Blood Cell Lysis

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.