The easiest way to analyze your samples by flow cytometry

Invitrogen Ready Flow reagents are ready-to-use solutions designed to allow you to stain your cells for analysis by flow cytometry with:

  • No calculations
  • No dilutions
  • No pipetting

Ready Flow reagents simplify the most common flow cytometric assays

Reach for the convenient dropper bottle, add 2 drops per 1 x 106 cells per milliliter of your single-cell suspension, and you're ready to incubate and analyze. Ready Flow reagents address your most common flow cytometry analysis needs, including dead cell identification and cell cycle (Figure 1) analysis. Preparing your samples for flow cytometry analysis is easy.

Figure 1. Cell cycle analysis with FxCycle Violet Ready Flow Reagent and Invitrogen Click-iT EdU Alexa Fluor 647 Flow Cytometry Assay Kit. Jurkat cells, a human T cell leukemia cell line, were pulsed with 10 µM EdU for 2 hours prior to detection with Alexa Fluor 647 azide. Cells were subsequently stained by adding 2 drops of FxCycle Violet Ready Flow Reagent and incubated for 30 minutes, at 25°C. Data was acquired on an Invitrogen Attune NxT Flow Cytometer using a 405 laser and 440/50 nm emission filter. Analysis of the population indicates the following distribution: apoptotic sub-G1 cells were 3.4%; G0/G1, 49.1%;  S 33.0%; and G2M 14.0%.

Ready Flow reagents

  SYTOX Green Ready Flow Reagent Propidium Iodide Ready Flow Reagent SYTOX AADvanced Ready Flow Reagent TO-PRO 3 Ready Flow Reagent
Readout Dead cells become highly fluorescent following labeling with reagent, allowing for easy distinction from live cells; no calculations, no dilutions, no pipetting.
Mechanism of labeling
Enters cells upon loss of membrane integrity and binds to nucleic acids.
Target Nucleic acids Nucleic acids Nucleic acids dsDNA
Fluorogenic >100-fold fluorescence enhancement upon nucleic acid binding Reagent is not fluorogenic; exhibits 20- to 30-fold fluorescence increase upon binding >500-fold fluorescence enhancement upon nucleic acid binding Reagent is not fluorogenic; exhibits fluorescence enhancement upon DNA binding
Storage
Room temperature Room temperature Room temperature Room temperature
Staining pattern fixable?
No No No No
Live-cell permeant? No No No No
Ex/Em of reagent (in nm) 504⁄523 535/617 546⁄647 642/661
Recommended excitation laser
  • 488 nm (blue)
  • 488 nm (blue)
  • 532 nm (green)
  • 561 nm (yellow)
  • 488 nm (blue)
  • 532 nm (green)
  • 561 nm (yellow)
  • 633 nm or 635 nm (red)
Recommended bandpass filter
  • 530/30
  • 574/26
  • 585/16
  • 590/40
  • 620/15
  • 695/40
  • 695/40
  • 670/10
Cat. No. R37168 R37169 R37173 R37170
  Hoechst 33342 Ready Flow Reagent FxCycle Violet Ready Flow Reagent Vybrant DyeCycle Violet Ready Flow Reagent
Readout

Fluorescent signal from reagent binding to DNA in live cells, allowing for cell cycle profiling and analysis; no calculations, no dilutions, no pipetting.

Fluorescent signal from reagent binding to DNA in fixed cells, allowing for cell cycle profiling and analysis; no calculations, no dilutions, no pipetting.

Fluorescent signal from reagent binding to DNA in live cells, allowing for cell cycle profiling and DNA content analysis; no calculations, no dilutions, no pipetting.

Target dsDNA dsDNA dsDNA
Mechanism of labeling The stain is a minor-groove DNA-binding reagent; binds preferentially to adenine–thymine regions of DNA. Stain is cell permeable and, after binding double-stranded DNA, emits a fluorescent signal that is proportional to the DNA mass.
Fluorogenic No. Fluorescence is greatly enhanced upon binding, but there is no modification of the dye structure.
Storage Room temperature Room temperature Room temperature
Live-cell assay? Yes No Yes
Staining pattern fixable? No No No
Fixed-cell assay? Yes Yes No
Ex/Em of reagent (in nm) 361/497 358/461 369/437
Recommended laser excitation UV (350 nm) or 405 nm UV (350 nm) or 405 nm UV (350 nm) or 405 nm
Recommended bandpass filter 440/50 450/50 440/40
Cat. No. R37165 R37166 R37172
  Annexin V
Pacific Blue
Ready Flow Reagent
Annexin V
Alexa Fluor 488
Ready Flow Reagent
Annexin V
Alexa Fluor 647
Ready Flow Reagent
Annexin V
APC
Ready Flow Reagent
CellEvent
Caspase-3/7
Green Ready
Flow Kit
Readout Fluorescent signal from reagent binding to phosphatidyl serine in apoptic cells; no calculations, no dilutions, no pipetting.  Fluorogenic substrate for detection of activated caspases 3 and 7 in apoptotic cells; no calculations, no dilutions, no pipetting. SYTOX AADvanced dead cell stain included in the kit for the detection of dead cells.
Mechanism of labeling In the presence of Ca2+, annexin V conjugates bind with high affinity to phosphatidylserine (PS), which becomes exposed on the outer leaflet of cells undergoing apoptosis. The annexin V conjugates can also pass through the compromised membranes of dead cells and bind to PS in the interior of the cell.  Recommend using a cell-impermeant dead cell stain in combination with annexin V conjugate staining to distinguish dead cells from apoptotic cells. The CellEvent reagent reagent consists of a four amino acid peptide (DEVD) conjugated to a nucleic acid binding dye. During apoptosis, caspase-3 and caspase-7 proteins are activated and are able to cleave the caspase 3/7 recognition sequence encoded in the DEVD peptide.
Target Binds to phosphatidylserine on apoptotic cell surfaces Caspase 3/7 proteins, dead cells
Fluorogenic No No No No Yes; cleavage of the recognition sequence and binding of DNA by the reagent labels apoptotic cells with a bright, signal
Storage Room temperature Room temperature Room temperature Room temperature Room
temperature
Staining pattern fixable No No No No No
Live-cell permeant No No No No Yes
Ex/Em of reagent (in nm) 410/455 495/519 650/665 650/660 CellEvent Caspase-3/7: ~511/533;
SYTOX AADvanced: 546/647
Recommended excitation laser 405 nm 488 nm 633 or 635 nm 633 or 635 nm CellEvent Caspase-3/7: 488 nm;
SYTOX AADvanced: 488 nm (blue), 532 nm (green) or 561 nm (yellow)
Recommended bandpass filter 450/50 530/30 661/8 661/8 CellEvent Caspase-3/7: 530/30;
SYTOX AADvanced: 695/40
Cat. No. R37177 R37174 R37175 R37176 R37167
  SYTOX Green Ready Flow Reagent Propidium Iodide Ready Flow Reagent SYTOX AADvanced Ready Flow Reagent TO-PRO 3 Ready Flow Reagent
Readout Dead cells become highly fluorescent following labeling with reagent, allowing for easy distinction from live cells; no calculations, no dilutions, no pipetting.
Mechanism of labeling
Enters cells upon loss of membrane integrity and binds to nucleic acids.
Target Nucleic acids Nucleic acids Nucleic acids dsDNA
Fluorogenic >100-fold fluorescence enhancement upon nucleic acid binding Reagent is not fluorogenic; exhibits 20- to 30-fold fluorescence increase upon binding >500-fold fluorescence enhancement upon nucleic acid binding Reagent is not fluorogenic; exhibits fluorescence enhancement upon DNA binding
Storage
Room temperature Room temperature Room temperature Room temperature
Staining pattern fixable?
No No No No
Live-cell permeant? No No No No
Ex/Em of reagent (in nm) 504⁄523 535/617 546⁄647 642/661
Recommended excitation laser
  • 488 nm (blue)
  • 488 nm (blue)
  • 532 nm (green)
  • 561 nm (yellow)
  • 488 nm (blue)
  • 532 nm (green)
  • 561 nm (yellow)
  • 633 nm or 635 nm (red)
Recommended bandpass filter
  • 530/30
  • 574/26
  • 585/16
  • 590/40
  • 620/15
  • 695/40
  • 695/40
  • 670/10
Cat. No. R37168 R37169 R37173 R37170
  Hoechst 33342 Ready Flow Reagent FxCycle Violet Ready Flow Reagent Vybrant DyeCycle Violet Ready Flow Reagent
Readout

Fluorescent signal from reagent binding to DNA in live cells, allowing for cell cycle profiling and analysis; no calculations, no dilutions, no pipetting.

Fluorescent signal from reagent binding to DNA in fixed cells, allowing for cell cycle profiling and analysis; no calculations, no dilutions, no pipetting.

Fluorescent signal from reagent binding to DNA in live cells, allowing for cell cycle profiling and DNA content analysis; no calculations, no dilutions, no pipetting.

Target dsDNA dsDNA dsDNA
Mechanism of labeling The stain is a minor-groove DNA-binding reagent; binds preferentially to adenine–thymine regions of DNA. Stain is cell permeable and, after binding double-stranded DNA, emits a fluorescent signal that is proportional to the DNA mass.
Fluorogenic No. Fluorescence is greatly enhanced upon binding, but there is no modification of the dye structure.
Storage Room temperature Room temperature Room temperature
Live-cell assay? Yes No Yes
Staining pattern fixable? No No No
Fixed-cell assay? Yes Yes No
Ex/Em of reagent (in nm) 361/497 358/461 369/437
Recommended laser excitation UV (350 nm) or 405 nm UV (350 nm) or 405 nm UV (350 nm) or 405 nm
Recommended bandpass filter 440/50 450/50 440/40
Cat. No. R37165 R37166 R37172
  Annexin V
Pacific Blue
Ready Flow Reagent
Annexin V
Alexa Fluor 488
Ready Flow Reagent
Annexin V
Alexa Fluor 647
Ready Flow Reagent
Annexin V
APC
Ready Flow Reagent
CellEvent
Caspase-3/7
Green Ready
Flow Kit
Readout Fluorescent signal from reagent binding to phosphatidyl serine in apoptic cells; no calculations, no dilutions, no pipetting.  Fluorogenic substrate for detection of activated caspases 3 and 7 in apoptotic cells; no calculations, no dilutions, no pipetting. SYTOX AADvanced dead cell stain included in the kit for the detection of dead cells.
Mechanism of labeling In the presence of Ca2+, annexin V conjugates bind with high affinity to phosphatidylserine (PS), which becomes exposed on the outer leaflet of cells undergoing apoptosis. The annexin V conjugates can also pass through the compromised membranes of dead cells and bind to PS in the interior of the cell.  Recommend using a cell-impermeant dead cell stain in combination with annexin V conjugate staining to distinguish dead cells from apoptotic cells. The CellEvent reagent reagent consists of a four amino acid peptide (DEVD) conjugated to a nucleic acid binding dye. During apoptosis, caspase-3 and caspase-7 proteins are activated and are able to cleave the caspase 3/7 recognition sequence encoded in the DEVD peptide.
Target Binds to phosphatidylserine on apoptotic cell surfaces Caspase 3/7 proteins, dead cells
Fluorogenic No No No No Yes; cleavage of the recognition sequence and binding of DNA by the reagent labels apoptotic cells with a bright, signal
Storage Room temperature Room temperature Room temperature Room temperature Room
temperature
Staining pattern fixable No No No No No
Live-cell permeant No No No No Yes
Ex/Em of reagent (in nm) 410/455 495/519 650/665 650/660 CellEvent Caspase-3/7: ~511/533;
SYTOX AADvanced: 546/647
Recommended excitation laser 405 nm 488 nm 633 or 635 nm 633 or 635 nm CellEvent Caspase-3/7: 488 nm;
SYTOX AADvanced: 488 nm (blue), 532 nm (green) or 561 nm (yellow)
Recommended bandpass filter 450/50 530/30 661/8 661/8 CellEvent Caspase-3/7: 530/30;
SYTOX AADvanced: 695/40
Cat. No. R37177 R37174 R37175 R37176 R37167

Resources

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