Cell Meter™ RX780 fixable viability dye

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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	1176.38
Solvent	DMSO

Spectral properties

Excitation (nm)	629
Emission (nm)	767

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	12171501

Overview SDS Protocol

Molecular weight

1176.38

Excitation (nm)

629

Emission (nm)

767

Discrimination and exclusion of dead cells from live cells allows cleaner separation and identification of cell populations. Cell Meter™ fixable viability dyes are a large family of cell-impermeable fluorescent viability dyes that are optimized to match the major excitation lasers of common flow cytometers, such as 350, 405, 488, 633 and 647 nm. These dyes are impermeant to live cells but permeant to cells with compromised membranes. They irreversibly react with amine- and thiol-containing proteins and other cellular components. Since dead or fixed cells with a compromised membrane more readily react with Cell Meter™ fixable cell stains, thus stain brighter than live cells with an intact membrane, these dyes can be used to assess live vs. dead status of mammalian cells. There are a few factors to be considered when using these dyes, e.g., the titration of each dye to ensure that live cells have minimal to no staining. Cell Meter™ RX780 fixable cell stain is optimized to be excited with the red lasers at 633/647 nm with emission at 780 nm. Compared to other commercially similar viability dyes, this fixable viability dye is much more robust and stable.

Platform

Flow cytometer

Excitation	633/647 nm laser
Emission	780 nm filter
Instrument specification(s)	APC-Cy7 channel

Fluorescence microscope

Excitation	Cy7 filter set
Emission	Cy7 filter set
Recommended plate	Black wall/clear bottom

Example protocol

AT A GLANCE

Protocol summary

Prepare samples in HHBS (0.5 mL/assay)
Add Cell Meter™ RX780 to the cell suspension
Stain the cells at room temperature for 20 - 60 minutes
Wash the cells
Fix the cells (optional)
Examine the sample with flow cytometer using 780 nm filter (APC-Cy7 channel) and/or fluorescence microscope using Cy7 filter set

Important

Thaw at room temperature before starting the experiment.

PREPARATION OF STOCK SOLUTIONS

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.

Cell Meter™ RX780 stock solution (500X)

Add 200 µL of DMSO into the Cell Meter™ RX780 vial to make 500X stock solution.
Note The unused Cell Meter™ RX780 stock solution should be divided into single use aliquots and stored at -20°C. Avoid repeated freeze/thaw cycles.

SAMPLE EXPERIMENTAL PROTOCOL

Treat cells as desired.
Wash cells once with HHBS or the azide- and serum/protein-free buffer of your choice.
Resuspend cells at 5 - 10 × 10⁶/mL in HHBS or in the azide- and serum/protein-free buffer of your choice.
Add 1 µL of 500X Cell Meter™ RX780 stock solution to 0.5 mL of cells/assay and mix it well.
Incubate at room temperature, 5% CO₂ incubator for 20 - 60 minutes, protected from light.
Note The optimal stain concentrations and incubation time should be experimentally determined for different cell lines.
Wash cells twice and resuspend cells with HHBS or the buffer of your choice.
Fix cells as desired (optional).
Analyze cells with a flow cytometer using 780 nm filter (APC-Cy7 channel) and/or fluorescence microscope using Cy7 filter set.

Calculators

Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of Cell Meter™ RX780 fixable viability dye 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	85.007 µL	425.033 µL	850.065 µL	4.25 mL	8.501 mL
5 mM	17.001 µL	85.007 µL	170.013 µL	850.065 µL	1.7 mL
10 mM	8.501 µL	42.503 µL	85.007 µL	425.033 µL	850.065 µL

Molarity calculator

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

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

Spectrum

Open in Advanced Spectrum Viewer

Spectral properties

Excitation (nm)	629
Emission (nm)	767

Product Family

Name	Excitation (nm)	Emission (nm)
Cell Meter™ BX520 fixable viability dye	491	516
Cell Meter™ BX590 fixable viability dye	492	579
Cell Meter™ BX650 fixable viability dye	518	654
Cell Meter™ IX830 fixable viability dye	811	822
Cell Meter™ RX660 fixable viability dye	649	664
Cell Meter™ RX700 fixable viability dye	690	713
Cell Meter™ VX450 fixable viability dye	406	445
Cell Meter™ VX500 fixable viability dye	433	498

Images

Figure 1. Detection of Jurkat cell viability by Cell Meter™ fixable viability dye. Jurkat cells were treated and stained with Cell Meter™ RX780 (Cat#22536), and then fixed in 3.7% formaldehyde and analyzed by flow cytometry. The dead cell population (Blue peak) is easily distinguished from the live cell population (Red peak) with APC-Cy7 channel, and nearly identical results were obtained before and after fixation

References

View all 50 references: Citation Explorer

PGC-1α Regulates Cell Proliferation and Invasion via AKT/GSK-3β/β-catenin Pathway in Human Colorectal Cancer SW620 and SW480 Cells.
Authors: Yun, Seong-Hoon and Park, Joo-In
Journal: Anticancer research (2020): 653-664

CFSE dilution to study human T and NK cell proliferation in vitro.
Authors: Terrén, Iñigo and Orrantia, Ane and Vitallé, Joana and Zenarruzabeitia, Olatz and Borrego, Francisco
Journal: Methods in enzymology (2020): 239-255

The miR-3940-5p inhibits cell proliferation of gingival mesenchymal stem cells.
Authors: Han, Xiao and Yang, Haoqing and Cao, Yangyang and Ge, Lihua and Han, Nannan and Zhang, Chen and Fan, Zhipeng and Yao, Rui
Journal: Oral diseases (2019): 1363-1373

Toll-Like Receptor 4 (TLR4)/Cyclooxygenase-2 (COX-2) Regulates Prostate Cancer Cell Proliferation, Migration, and Invasion by NF-κB Activation.
Authors: Wang, Wei and Wang, Jiye
Journal: Medical science monitor : international medical journal of experimental and clinical research (2018): 5588-5597

Evaluation of Cell Proliferation and Apoptosis in Immunotoxicity Testing.
Authors: Nagarkatti, Mitzi and Rieder, Sadiye Amcaoglu and Nagarkatti, Prakash S
Journal: Methods in molecular biology (Clifton, N.J.) (2018): 209-230

Quantitative analysis of cell proliferation by a dye dilution assay: Application to cell lines and cocultures.
Authors: Chung, Soobin and Kim, Seol-Hee and Seo, Yuri and Kim, Sook-Kyung and Lee, Ji Youn
Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology (2017): 704-712

[Myeloid-derived suppressor cells promoted autologous B cell proliferation in rheumatoid arthritis].
Authors: Chen, W and Hu, F L and Liu, H J and Xu, L L and Li, Y N and Li, Z G
Journal: Beijing da xue xue bao. Yi xue ban = Journal of Peking University. Health sciences (2017): 819-823

Gag-Specific CD4 T Cell Proliferation, Plasmacytoid Dendritic Cells, and Ethnicity in Perinatally HIV-1-Infected Youths: The ANRS-EP38-IMMIP Study.
Authors: Scott-Algara, Daniel and Warszawski, Josiane and Chenadec, Jérôme Le and Didier, Céline and Montange, Thomas and Viard, Jean-Paul and Dollfus, Catherine and Avettand-Fenoel, Véronique and Rouzioux, Christine and Blanche, Stéphane and Buseyne, Florence
Journal: AIDS research and human retroviruses (2017): 21-28

Hypoxia inhibits mesenchymal stem cell proliferation through HIF1α-dependent regulation of P27.
Authors: Kumar, Sanjay and Vaidya, Meenal
Journal: Molecular and cellular biochemistry (2016): 29-38

Vanadium pentoxide prevents NK-92MI cell proliferation and IFNγ secretion through sustained JAK3 phosphorylation.
Authors: Gallardo-Vera, Francisco and Diaz, Daniel and Tapia-Rodriguez, Miguel and Fortoul van der Goes, Teresa and Masso, Felipe and Rendon-Huerta, Erika and Montaño, Luis F
Journal: Journal of immunotoxicology (2016): 27-37