Ready Flow Ready-to-Use Flow Cytometry Reagents

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.

FxCycle Violet Ready Flow Reagent

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 GreenPropidium IodideSYTOX AADvancedTO-PRO 3DRAQ7
ReadoutDead 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.
TargetNucleic acidsNucleic acidsNucleic acidsdsDNAdsDNA
Fluorogenic>100-fold fluorescence enhancement upon nucleic acid bindingReagent is not fluorogenic; exhibits 20- to 30-fold fluorescence increase upon binding>500-fold fluorescence enhancement upon nucleic acid bindingReagent is not fluorogenic; exhibits fluorescence enhancement upon DNA bindingReagent is not fluorogenic
StorageRoom temperatureRoom temperatureRoom temperatureRoom temperatureRoom temperature
Staining pattern fixable?
NoNoNoNoNo
Live-cell permeant?NoNoNoNoNo
Ex/Em of reagent (in nm)504/523535/617546/647642/661488 to 647/677
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)
  • 633 nm (red)
Recommended bandpass filter
  • 530/30
  • 574/26
  • 585/16
  • 590/40
  • 620/15
  • 695/40
  • 695/40
  • 670/10
  • 695 LP
  • 715 LP
  • 780 LP
Cat. No.R37168R37169R37173R37170D15107
 Hoechst 33342 Ready Flow ReagentFxCycle Violet Ready Flow ReagentVybrant 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.

TargetdsDNAdsDNAdsDNA
Mechanism of labelingThe 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.
FluorogenicNo. Fluorescence is greatly enhanced upon binding, but there is no modification of the dye structure.
StorageRoom temperatureRoom temperatureRoom temperature
Live-cell assay?YesNoYes
Staining pattern fixable?NoNoNo
Fixed-cell assay?YesYesNo
Ex/Em of reagent (in nm)361/497358/461369/437
Recommended laser excitationUV (350 nm) or 405 nmUV (350 nm) or 405 nmUV (350 nm) or 405 nm
Recommended bandpass filter440/50450/50440/40
Cat. No.R37165R37166R37172
 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
ReadoutFluorescent 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 labelingIn 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.
TargetBinds to phosphatidylserine on apoptotic cell surfacesCaspase 3/7 proteins, dead cells
FluorogenicNoNoNoNoYes; cleavage of the recognition sequence and binding of DNA by the reagent labels apoptotic cells with a bright, signal
StorageRoom temperatureRoom temperatureRoom temperatureRoom temperatureRoom
temperature
Staining pattern fixableNoNoNoNoNo
Live-cell permeantNoNoNoNoYes
Ex/Em of reagent (in nm)410/455495/519650/665650/660CellEvent Caspase-3/7: ~511/533;
SYTOX AADvanced: 546/647
Recommended excitation laser405 nm488 nm633 or 635 nm633 or 635 nmCellEvent Caspase-3/7: 488 nm;
SYTOX AADvanced: 488 nm (blue), 532 nm (green) or 561 nm (yellow)
Recommended bandpass filter450/50530/30661/8661/8CellEvent Caspase-3/7: 530/30;
SYTOX AADvanced: 695/40
Cat. No.R37177R37174R37175R37176R37167
 SYTOX GreenPropidium IodideSYTOX AADvancedTO-PRO 3DRAQ7
ReadoutDead 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.
TargetNucleic acidsNucleic acidsNucleic acidsdsDNAdsDNA
Fluorogenic>100-fold fluorescence enhancement upon nucleic acid bindingReagent is not fluorogenic; exhibits 20- to 30-fold fluorescence increase upon binding>500-fold fluorescence enhancement upon nucleic acid bindingReagent is not fluorogenic; exhibits fluorescence enhancement upon DNA bindingReagent is not fluorogenic
StorageRoom temperatureRoom temperatureRoom temperatureRoom temperatureRoom temperature
Staining pattern fixable?
NoNoNoNoNo
Live-cell permeant?NoNoNoNoNo
Ex/Em of reagent (in nm)504/523535/617546/647642/661488 to 647/677
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)
  • 633 nm (red)
Recommended bandpass filter
  • 530/30
  • 574/26
  • 585/16
  • 590/40
  • 620/15
  • 695/40
  • 695/40
  • 670/10
  • 695 LP
  • 715 LP
  • 780 LP
Cat. No.R37168R37169R37173R37170D15107
 Hoechst 33342 Ready Flow ReagentFxCycle Violet Ready Flow ReagentVybrant 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.

TargetdsDNAdsDNAdsDNA
Mechanism of labelingThe 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.
FluorogenicNo. Fluorescence is greatly enhanced upon binding, but there is no modification of the dye structure.
StorageRoom temperatureRoom temperatureRoom temperature
Live-cell assay?YesNoYes
Staining pattern fixable?NoNoNo
Fixed-cell assay?YesYesNo
Ex/Em of reagent (in nm)361/497358/461369/437
Recommended laser excitationUV (350 nm) or 405 nmUV (350 nm) or 405 nmUV (350 nm) or 405 nm
Recommended bandpass filter440/50450/50440/40
Cat. No.R37165R37166R37172
 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
ReadoutFluorescent 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 labelingIn 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.
TargetBinds to phosphatidylserine on apoptotic cell surfacesCaspase 3/7 proteins, dead cells
FluorogenicNoNoNoNoYes; cleavage of the recognition sequence and binding of DNA by the reagent labels apoptotic cells with a bright, signal
StorageRoom temperatureRoom temperatureRoom temperatureRoom temperatureRoom
temperature
Staining pattern fixableNoNoNoNoNo
Live-cell permeantNoNoNoNoYes
Ex/Em of reagent (in nm)410/455495/519650/665650/660CellEvent Caspase-3/7: ~511/533;
SYTOX AADvanced: 546/647
Recommended excitation laser405 nm488 nm633 or 635 nm633 or 635 nmCellEvent Caspase-3/7: 488 nm;
SYTOX AADvanced: 488 nm (blue), 532 nm (green) or 561 nm (yellow)
Recommended bandpass filter450/50530/30661/8661/8CellEvent Caspase-3/7: 530/30;
SYTOX AADvanced: 695/40
Cat. No.R37177R37174R37175R37176R37167

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Not for resale. Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.