CytoTrace™ Ultra Green

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Additional ordering information

Telephone	1-800-990-8053
Fax	1-800-609-2943
Email	sales@aatbio.com
International	See distributors
Bulk request	Inquire
Custom size	Inquire
Shipping	Standard overnight for United States, inquire for international

Physical properties

Molecular weight	822.72
Solvent	DMSO

Storage, safety and handling

H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
Storage	Freeze (< -15 °C); Minimize light exposure
UNSPSC	12352200

Overview SDS Protocol

Example protocol

AT A GLANCE

Protocol summary

Prepare cells with test compounds
Add 0.5 to 5 µM CytoTrace™ Ultra Green working solution
Incubate dyes with cells at room temperature or 37 ^oC for 15 to 30 minutes
Remove the dye working solution
Analyze with a flow cytometer with Ex/Em = 490/520 nm-FITC filter set

PREPARATION OF STOCK SOLUTION

Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles.

CytoTrace™ Ultra Green stock solution (2-10 mM):
Add appropriate amount of DMSO and mix well to make CytoTrace™ Ultra Green stock solution (2-10 mM). Note: The stock solution should be used promptly; any remaining solution should be aliquoted and frozen at < -20 ^oC. Avoid repeated freeze-thaw cycles, and protect from light.

PREPARATION OF WORKING SOLUTION

CytoTrace™ Ultra Green working solution:
Prepare a 0.5 to 5 µM dye working solution right before use by diluting the DMSO stock solution from with Hanks and 20 mM Hepes buffer (HHBS) or the buffer of your choice, pH 7. Mix them well by vortexing. Note: In some cell types, lower concentration may be required to stain the cells. We recommend optimizing optimal concentration for each cell type before performing experiment.

SAMPLE EXPERIMENTAL PROTOCOL

Treat cells with test compounds for a desired period of time.
Centrifuge the cells to get 2-10 × 10⁵cells per tube.
Resuspend cells in 500 µL of the CytoTrace™ Ultra Green working solution.
Incubate cells with a dye solution at room temperature or 37 °C for 15 to 30 min, protected from light.
Remove the dye working solution from the cells; Optional: The cells can be fixed with 4% formaldehyde.
Wash the cells with HHBS or buffer of your choice once.
Resuspend cells in 500 µL of pre-warmed HHBS or medium to get 2-10 × 10⁵cells per tube.
Monitor the fluorescence change at Ex/Em = 490/520 nm with a flow cytometer or a fluorescence microscope with FITC filter set.

Product #	Indicator	Size	Molecular Weight	Ex/Em (nm)	Solvent
22014	CytoTrace™ Orange CMTMR	10x50 mg	554.04	541/565	DMSO
22015	CytoTrace™ Red CMPTX	10x50 mg	686.25	577/602	DMSO
22016	CytoTrace™ Red CFDA	1 mg	652.43	560/574	DMSO
22017	CytoTrace™ Green CMFDA	1 mg	464.86	494/521	DMSO
21800	CytoTrace™ Ultra Green	1 mg	822.72	494/521	DMSO
22020	FDA (Fluorescein diacetate)	1 g	416.83	494/521	DMSO

Calculators

Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of CytoTrace™ Ultra Green to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

	0.1 mg	0.5 mg	1 mg	5 mg	10 mg
1 mM	121.548 µL	607.74 µL	1.215 mL	6.077 mL	12.155 mL
5 mM	24.31 µL	121.548 µL	243.096 µL	1.215 mL	2.431 mL
10 mM	12.155 µL	60.774 µL	121.548 µL	607.74 µL	1.215 mL

Molarity calculator

Enter any two values (mass, volume, concentration) to calculate the third.

Mass (Calculate)		Molecular weight		Volume (Calculate)		Concentration (Calculate)		Moles
	/		=		x		=

Product Family

Name	Excitation (nm)	Emission (nm)
CytoTrace™ Green CMFDA	498	517

Images

Figure 1. The comparison in the fluorescence intensity of CytoTrace™ Ultra Green with CMFDA in Jurkat cells. Jurkat cells were dye loaded with CytoTrace™ Ultra Green or CMFDA for 30 minutes in a 37 ^oC, 5% CO₂ incubator. The fluorescence intensity was measured using ACEA NovoCyte 3000 flow cytometer with FITC channel.

Citations

View all 5 citations: Citation Explorer

SPF-CellTracker: Tracking Multiple Cells with Strongly-Correlated Moves Using a Spatial Particle Filter
Authors: Hirose, O., Kawaguchi, S., Tokunaga, T., Toyoshima, Y., Teramoto, T., Kuge, S., Ishihara, T., Iino, Y., Yoshida, R.
Journal: IEEE/ACM Trans Comput Biol Bioinform (2018): 1822-1831

Challenges in using CellTracker Green on foraminifers that host algal endosymbionts
Authors: Ross, B. J., Hallock, P.
Journal: PeerJ (2018): e5304

CellTracker (not only) for dummies
Authors: Piccinini, F., Kiss, A., Horvath, P.
Journal: Bioinformatics (2016): 955-7

In vitro evaluation of the effects of cysticidal drugs in the Taenia crassiceps cysticerci ORF strain using the fluorescent CellTracker CMFDA
Authors: Trejo-Chavez, H., Garcia-Vilchis, D., Reynoso-Ducoing, O., Ambrosio, J. R.
Journal: Exp Parasitol (2011): 294-9

Survival of benthic foraminifera under hypoxic conditions: results of an experimental study using the CellTracker Green method
Authors: Pucci, F., Geslin, E., Barras, C., Morigi, C., Sabbatini, A., Negri, A., Jorissen, F. J.
Journal: Mar Pollut Bull (2009): 336-51

References

View all 23 references: Citation Explorer

Fluorescence-Based Transport Assays Revisited in a Human Renal Proximal Tubule Cell Line
Authors: Caetano-Pinto P, Janssen MJ, Gijzen L, Verscheijden L, Wilmer MJ, Masereeuw R.
Journal: Mol Pharm (2016): 933

The variable chemotherapeutic response of Malabaricone-A in leukemic and solid tumor cell lines depends on the degree of redox imbalance
Authors: Manna A, De Sarkar S, De S, Bauri AK, Chattopadhyay S, Chatterjee M.
Journal: Phytomedicine (2015): 713

Cell membrane tracker based on restriction of intramolecular rotation
Authors: Zhang C, Jin S, Yang K, Xue X, Li Z, Jiang Y, Chen WQ, Dai L, Zou G, Liang XJ.
Journal: ACS Appl Mater Interfaces (2014): 8971

Evaluation of stability and sensitivity of cell fluorescent labels when used for cell migration
Authors: Beem E, Segal MS.
Journal: J Fluoresc (2013): 975

A multiple model probability hypothesis density tracker for time-lapse cell microscopy sequences
Authors: Rezatofighi SH, Gould S, Vo BN, Mele K, Hughes WE, Hartley R.
Journal: Inf Process Med Imaging (2013): 110

TLM-Tracker: software for cell segmentation, tracking and lineage analysis in time-lapse microscopy movies
Authors: Klein J, Leupold S, Biegler I, Biedendieck R, Munch R, Jahn D.
Journal: Bioinformatics (2012): 2276

The interplay between Leishmania promastigotes and human Natural Killer cells in vitro leads to direct lysis of Leishmania by NK cells and modulation of NK cell activity by Leishmania promastigotes
Authors: Lieke T, Nylen S, Eidsmo L, Schmetz C, Berg L, Akuffo H.
Journal: Parasitology (2011): 1898

The exocytosis of fluorescent nanodiamond and its use as a long-term cell tracker
Authors: Fang CY, Vaijayanthimala V, Cheng CA, Yeh SH, Chang CF, Li CL, Chang HC.
Journal: Small (2011): 3363

Horizontal DNA transfer from donor to host cells as an alternative mechanism of epithelial chimerism after allogeneic hematopoietic cell transplantation
Authors: Waterhouse M, Themeli M, Bertz H, Zoumbos N, Finke J, Spyridonidis A.
Journal: Biol Blood Marrow Transplant (2011): 319

Overlap-Based Cell Tracker
Authors: Chalfoun J, Cardone A, Dima AA, Allen DP, Halter MW.
Journal: J Res Natl Inst Stand Technol (2010): 477