Ready-to-Use Flow Cytometry Reagents | Thermo Fisher Scientific

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.

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/G_1, 49.1%; S 33.0%; and G₂M 14.0%.

Ready Flow reagents

	SYTOX Green	Propidium Iodide	SYTOX AADvanced	TO-PRO 3	DRAQ7
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	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	Reagent is not fluorogenic
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	No
Ex/Em of reagent (in nm)	504/523	535/617	546/647	642/661	488 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.	R37168	R37169	R37173	R37170	D15107

	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 apoptotic 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

Viability

	SYTOX Green	Propidium Iodide	SYTOX AADvanced	TO-PRO 3	DRAQ7
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	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	Reagent is not fluorogenic
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	No
Ex/Em of reagent (in nm)	504/523	535/617	546/647	642/661	488 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.	R37168	R37169	R37173	R37170	D15107

Cell cycle

	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

Apoptosis

	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 apoptotic 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

Fluorophore and reagent selection guide for flow cytometry

Download Flow Cytometry Protocols Handbook

Spectral Flow Cytometry Fundamentals

Invitrogen eBioscience Resources—Selection guides, Best Protocols, product performance and more.

Intracellular Staining for Flow Cytometry How-To Video—for detecting cytokines and intranuclear markers.

Flow Cytometry Learning Center—Access flow cytometry educational resources for better experiment planning and execution.

Flow Cytometry Panel Builder—Design your flow cytometry panel with this online tool for a simplified, customizable experience to fit your needs.

5 Steps Resources

See all 5 Steps Workflows

Support

Flow Cytometry Support Center—Find technical support recommendations for your flow cytometry workflows, including tips for experimental setup and in-depth troubleshooting help.

Flow Cytometry Panel Design Support—Work with one of our technical sales specialists to discuss your experimental needs and guide you through the process.

Flow Cytometry Protocols

Flow Cytometry Application Notes and Posters