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While some assays utilize antibodies to study cell health, proliferation, cell cycle or apoptosis, other types of experiments can rely on non-antibody based methods of assessment, often called non-antibody chemical probes. These are reagents that localize to an organelle or indicate health based on chemical characteristics like hydrophobicity, charge, size, and enzymatic activation.

 

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Image showing a cell, the organelles it contains within, and the reagents BioLegend provides to investigate them.

We have divided our chemical probes into tables based on their function or localization to help you choose the right reagent for your labeling application. Cell type suitability indicates whether this reagent labels live cells exclusively or whether it can also label the dead cells in a live cell culture or cells that have been previously fixed with buffers like paraformaldehyde or methanol. Sample suitability indicates whether this reagent can be used to label already fixed tissue sections or single cells in suspension or cell culture. Application indicates their suitability for being analyzed by flow cytometry, microscopy platforms or both. Fixation simply indicates whether the reagent, once used to label a cell sample, will be retained with a paraformaldehyde fixation.

 

Expand tables below to view chemical probes by localization and/or function.

 

  Spectra Functionality Cell Type Suitability Sample Suitability Application Fixation
Chemical Probe     Live Dead/ Fixed Tissue Cell Culture/ Single Cells Flow Cytometry Microscopy Retention with PFA Treatment
Apotracker™ Green View Spectra Apoptosis indicator    
Apotracker™ Tetra Alexa Fluor® 647 View Spectra Apoptosis indicator    
Annexin V Several formats available Apoptosis indicator      
7-AAD View Spectra Dead cell indicator    
Zombie UV387™ View Spectra Fixable cell death indicator    
Zombie UV™ View Spectra Fixable cell death indicator    
Zombie Violet™ View Spectra Fixable cell death indicator    
Zombie Aqua™ View Spectra Fixable cell death indicator    
Zombie Yellow™ View Spectra Fixable cell death indicator    
Zombie Green™ View Spectra Fixable cell death indicator    
Zombie B550™ View Spectra Fixable cell death indicator    
Zombie YG581™ View Spectra Fixable cell death indicator    
Zombie Red™ View Spectra Fixable cell death indicator    
Zombie R685™ View Spectra Fixable cell death indicator    
Zombie R718™ View Spectra Fixable cell death indicator    
Zombie NIR™ View Spectra Fixable cell death indicator    
Helix NP™ Blue View Spectra Viability indicator    
Helix NP™ Green View Spectra Viability indicator    
Helix NP™ NIR View Spectra Viability indicator    

 

 

  Spectra Functionality Cell Type Suitability Sample Suitability Application Fixation
Chemical Probe     Live Dead/ Fixed Tissue Cell Culture/ Single Cells Flow Cytometry Microscopy Retention with PFA Treatment
Flash Phalloidin™ Green 488 View Spectra F-actin staining     N/A
Flash Phalloidin™ Red 594 View Spectra F-actin staining     N/A

 

(+)    Cytoplasm

 

  Spectra Functionality Cell Type Suitability Sample Suitability Application Fixation
Chemical Probe     Live Dead/ Fixed Tissue Cell Culture/ Single Cells Flow Cytometry Microscopy Retention with PFA Treatment
Calcein-AM View Spectra Vitality indicator      
Calcein Red-AM View Spectra Vitality indicator      
Calcein Violet-AM View Spectra Vitality indicator      
Cellular Senescence Kit Red Exmax/Emmax 544, 567 nm Cellular senescence/cell cycle    
CFSE View Spectra Proliferation, cell tracking  
Tag-it Violet™ View Spectra Proliferation, cell tracking  

 

 

  Spectra Functionality Cell Type Suitability Sample Suitability Application Fixation
Chemical Probe     Live Dead/ Fixed Tissue Cell Culture/ Single Cells Flow Cytometry Microscopy Retention with PFA Treatment
Endoplasmic Reticulum Probe Green View Spectra Organelle staining      
Endoplasmic Reticulum Probe Red View Spectra Organelle staining      

 

(+)    Golgi Appartus

 

  Spectra Functionality Cell Type Suitability Sample Suitability Application Fixation
Chemical Probe     Live Dead/ Fixed Tissue Cell Culture/ Single Cells Flow Cytometry Microscopy Retention with PFA Treatment
Golgi Detection Probe Red Exmax/Emmax 
544, 570 nm
Organelle staining        

 

(+)    Lysosome

 

  Spectra Functionality Cell Type Suitability Sample Suitability Application Fixation
Chemical Probe     Live Dead/ Fixed Tissue Cell Culture/ Single Cells Flow Cytometry Microscopy Retention with PFA Treatment
Lysosome Probe Green View Spectra Organelle staining      
Lysosome Probe Orange View Spectra Organelle staining      
Lysosome Probe Red View Spectra Organelle staining      
Lysosome Probe Deep Red View Spectra Organelle staining      
Lysosome Probe NIR View Spectra Organelle staining      

 

(+)    Mitochondria

 

  Spectra Functionality Cell Type Suitability Sample Suitability Application Fixation
Chemical Probe     Live Dead/ Fixed Tissue Cell Culture/ Single Cells Flow Cytometry Microscopy Retention with PFA Treatment
JC-10 Mitochondrial Membrane Potential Kit View Spectra Depolarization/apoptosis indicator      
MitoSpy™ Green FM View Spectra Vitality indicator      
MitoSpy™ Orange CMTMRos View Spectra Vitality indicator    
MitoSpy™ Red CMXRos View Spectra Vitality indicator    
MitoSpy™ NIR DiIC1(5) View Spectra Vitality indicator      

 

(+)    Nucleus

 

  Spectra Functionality Cell Type Suitability Sample Suitability Application Fixation
Chemical Probe     Live Dead/ Fixed Tissue Cell Culture/ Single Cells Flow Cytometry Microscopy Retention with PFA Treatment
Helix NP™ Blue View Spectra Viability indicator    
Helix NP™ Green View Spectra Viability indicator    
Helix NP™ NIR View Spectra Viability indicator    
CytoPhase™ Violet View Spectra Cell cycle/viability indicator  
DRAQ5™ View Spectra Cell cycle/viability indicator  
DRAQ7™ View Spectra Cell cycle/viability indicator    
Propidium Iodide View Spectra Cell cycle/viability indicator    
DAPI View Spectra Nuclear stain/cell cycle    

 

 

  Spectra Functionality Cell Type Suitability Sample Suitability Application Fixation
Chemical Probe     Live Dead/ Fixed Tissue Cell Culture/ Single Cells Flow Cytometry Microscopy Retention with PFA Treatment
Autophagy Detection Probe Blue Exmax/Emmax 
330, 520 nm
Autophagy indicator        
Autophagy Detection Probe Green Exmax/Emmax 
485, 530 nm
Autophagy indicator        
Phagocytosis Detection Kit Red Exmax/Emmax 
570, 600 nm
Phagocytosis activity      

 

Cellular division for any cell type is dependent on the inherent function, location and the response of cells to repair, apoptosis, or death. To divide, cells must duplicate a copy of their DNA, increase mitochondrial density, and assemble/synthesize microtubules during interphase. G2 is a checkpoint stage of interphase where the cell has two sets of dsDNA and must commit to mitosis. Mitosis is the actual division stage where two daughter cells are created. There can be symmetrical or asymmetrical division depending on the cell and tissue type. Mitosis can be further subdivided into prophase, metaphase, anaphase, and telophase as the nucleus undergoes division and chromatids are pulled away from one another. After mitosis, the cell will undergo the G0/G1 phase when the cells rest to become ready for the next round of replication.

cycle

During G0/G1, S, and G2 phases, some fluorogenic nucleic acid stains selectively bind dsDNA stoichiometrically to give us measurements of DNA mass based on fluorescence intensity. Cell-permeant nucleic acid stains can be used in instances where cells will be analyzed live. Propidium IodideDAPIDRAQ5™DRAQ7™CytoPhase™ VioletHelix NP™ NIR,  Helix NP™ Blue, and Helix NP™ Green can all be used to stain fixed cells for cell cycle analysis. However, only CytoPhase™ Violet and DRAQ5™ can be used to assess DNA content of live cells.

 

For additional reagents (including antibodies) on cell cycle analysis or DNA replication, check here.

Live and Dead Cell Cycle Analysis

CytoPhase™ Violet

 

Live Ramos cells treated with 5 µM CytoPhase™ Violet dye for 90 minutes at 37°C. 

DRAQ5™

 

Live C57BL/6 mouse bone marrow cells were stained with DRAQ5™.
 

Helix NP™ NIR

 

C57BL/6 mouse thymus cells were fixed using 70% chilled ethanol. The cells were incubated for one hour at -20°C, washed, then stained with Helix NP™ NIR at 5 µM.

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