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SYTOX Green

SYTOX Green is a fluorescent dye used in various biological applications, particularly for detecting cell death and nucleic acid staining.
It is a cell-impermeable dye that only enters cells with compromised membranes, making it a useful tool for identifying dead or dying cells.
SYTOX Green binds to DNA, resulting in a bright green fluorescent signal that can be detected using fluorescence microscopy or flow cytometry.
Researchers can leverage SYTOX Green to optimize their experimental protocols, enhance reproducibility, and gain accurate insights into their samples.
PubCompare.ai, an AI-driven platform, can help scientists easily locate relevant SYTOX Green protocols from literature, preprints, and patents, and use advanced comparisons to identify the best products and methods for their specific research needs.
This can help improve the accuracy and reliability of SYTOX Green-based experiments and lead to more robust, reproducible results.

Most cited protocols related to «SYTOX Green»

Maintenance and Differentiation of Cell Lines

Cited 26 times

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Publication 2014

benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone Biological Factors BMS345541 Bone Marrow Cells CASP8 protein, human Culture Media, Conditioned Cycloheximide Femur Fetal Bovine Serum Glucose Glutamine HT29 Cells Macrophage Colony-Stimulating Factor necrostatin-1 Neutrophil NIH 3T3 Cells Penicillins Poly I-C Promega quinoline-val-asp(OMe)-CH2-OPH RNA, Small Interfering Streptomycin Sulfoxide, Dimethyl SYTOX Green Tibia Tumor Necrosis Factor-alpha

Immunofluorescent Labeling of Autophagy Markers in Drosophila

Cited 18 times

LTR stainings were performed by incubating dissected L3 stage fat bodies in 100 nM LTR (Invitrogen) for 5 min. For immunofluorescent labeling, bisected larvae were fixed overnight in 3.7% paraformaldehyde at 4°C and blocked in PBS with 0.1% Triton X-100, 0.05% sodium deoxycholate, and 3% goat serum for 3 h followed by overnight incubations with primary and secondary antibodies in blocking buffer at 4°C (Juhász et al., 2008 (link); Pircs et al., 2012 (link)). We used chicken anti-GFP (1:1,500; Invitrogen), rabbit anti-Atg5 (1:100; Sigma-Aldrich), rabbit anti-Atg8a (1:500; provided by K. Kohler, Eidgenössische Technische Hochschule Zürich, Zurich, Switzerland; Barth et al., 2011 (link)), rabbit anti-p62 (1:2,000; Pircs et al., 2012 (link)), rabbit anti–active caspase 3 (1:300; Cell Signaling Technology), rat anti-Atg8a (1:300), and rat anti-Syx17 (1:300; this study) primary and Alexa Fluor 488 anti–chicken, Alexa Fluor 488 anti–rabbit, Alexa Fluor 568 anti–rat, and Alexa Fluor 647 anti–rabbit (all 1:1,500; Invitrogen) secondary antibodies. For TUNEL stainings, adult heads and half-thoraces were fixed in 3.7% paraformaldehyde overnight at 4°C and embedded into paraffin following standard protocols. Sections were processed using In Situ Cell Death Detection Kit, tetramethylrhodamine red (Roche) with SYTOX green DNA stain (Juhász et al., 2007 (link)). Images were obtained on a microscope (Axio Imager.M2; Carl Zeiss) equipped with a grid confocal unit (ApoTome.2; Carl Zeiss) at room temperature, using Plan-Neofluar 20×, 0.5 NA (air), 40×, 0.75 NA (air), and 100×, 1.3 NA (oil) objectives, a camera (AxioCam MRm; Carl Zeiss), and AxioVision software (Carl Zeiss). Microscope settings were identical for experiments of the same kind. Primary images were processed in AxioVision and Photoshop (Adobe) to produce final figures. Note that Alexa Fluor 568 or 647 channels are pseudocolored magenta.

Takáts S., Nagy P., Varga Á., Pircs K., Kárpáti M., Varga K., Kovács A.L., Hegedűs K, & Juhász G. (2013). Autophagosomal Syntaxin17-dependent lysosomal degradation maintains neuronal function in Drosophila. The Journal of Cell Biology, 201(4), 531-539.

Publication 2013

Adult alexa 568 alexa fluor 488 Alexa Fluor 647 Antibodies Antibodies, Blocking Buffers Caspase 3 Cell Death Chest Chickens Deoxycholic Acid, Monosodium Salt Fat Body Fluorescent Antibody Technique Goat Head In Situ Nick-End Labeling Larva Microscopy Paraffin Embedding paraform Rabbits Rosaniline Dyes Serum Staining Stains SYTOX Green tetramethylrhodamine Triton X-100

Cytotoxicity Assay for Breast Cancer

Cited 8 times

Breast cancer cells and MCF-10A cells seeded at 1 × 10⁴ per well in 96-well plates were treated with Mito-ChM or Mito-ChMAc for 24 h, and dead cells were monitored in the presence of 200 nM Sytox Green (Invitrogen). The Sytox method labels the nuclei of dead cells yielding green fluorescence. Fluorescence intensities from the dead cells in 96-well plate were acquired in real time every 5 min for first 4 h, then every 15 min after 4 h using a plate reader (BMG Labtech, Inc.) equipped with atmosphere controller set at 37°C and 5% CO2:95% air using a fluorescence detection with 485 nm excitation and 535 nm emission. To measure the total cell number, all of the samples in each treatment group were permeabilized by adding Triton X 100 (0.065%) in the presence of Sytox Green for 3 h, and maximal fluorescence intensities were taken as 100%. Data are represented as a percentage of dead cells after normalization to total cell number for each group.
The IncuCyte™ Live-Cell Imaging system was used for kinetic monitoring of cytotoxicity as determined by Sytox Green staining at regular cell culture condition [23 (link)]. Additionally, phase-contrast and fluorescent images were automatically collected for each time point to determine morphological cell changes.
For clonogenic assay, MCF-7, MDA-MB-231 and MCF-10A cells were seeded at 300 cells per dish in 6 cm diameter cell culture dishes and treated with Mito-ChM for 4 h. After 7–14 days, the number of colonies formed was determined. The cell survival fractions were calculated according to a published protocol [24 (link)].

Cheng G., Zielonka J., McAllister D.M., Mackinnon AC J.r., Joseph J., Dwinell M.B, & Kalyanaraman B. (2013). Mitochondria-targeted vitamin E analogs inhibit breast cancer cell energy metabolism and promote cell death. BMC Cancer, 13, 285.

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Publication 2013

Atmosphere Biological Assay Breast Carcinoma Cell Culture Techniques Cell Nucleus Cells Cell Survival Cytotoxin Fluorescence Hyperostosis, Diffuse Idiopathic Skeletal Kinetics Microscopy, Phase-Contrast Mitomycin SYTOX Green Triton X-100

Embryo and Seed Staining Protocols

Cited 8 times

For staining of embryos, seeds were imbibed over night in water before being dissected out of their seed coats with fine needles. Fixation of tissue was performed over night at 4°C in a solution of 4% paraformaldehyde in 100 mM sodium phosphate buffer pH7. Propidium iodide staining: propidium iodide (Molecular Probes, Eugene, USA) was used as a 10 μg/ml solution in water. Plants were stained for 5 minutes. SYTOX staining: SYTOX green, orange, and blue were supplied as solution in DMSO (Molecular Probes, Eugene, USA) and were diluted to 250 nM in water. Staining times are indicated in the text. Fluorescein diacetate staining: Fluorescein diacetate (Sigma-Aldrich, Saint Louis, USA) was used as a 5 μg/ml solution in water. Plants were stained for 20 to 30 minutes. All images were taken within the first 30 minutes after staining.

Truernit E, & Haseloff J. (2008). A simple way to identify non-viable cells within living plant tissue using confocal microscopy. Plant Methods, 4, 15.

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Publication 2008

Buffers diacetylfluorescein Embryo Molecular Probes Needles paraform Plants Propidium Iodide sodium phosphate Sulfoxide, Dimethyl SYTOX Green Tissue Fixation

Visualizing Neutrophil Extracellular Traps

Cited 7 times

Visualization of NETs by immunofluorescence was performed using an adapted version of the protocol described by Brinkmann et al. (Brinkmann et al., 2010 (link)). Briefly, resting or primed neutrophils (2 × 10⁵ in assay media) were seeded onto 13-mm circular glass coverslips (VWR International, Leicestershire, UK) and incubated for 30 min at 37°C in a 5% CO₂ atmosphere to allow for cell adherence. Postincubation, neutrophils were stimulated with PMA (25 nm), IL-8 (10 ng mL⁻¹), LPS (100 ng mL⁻¹) or assay media for 3 h (37°C, 5% CO₂), after which cells were fixed in 2% paraformaldehyde for 30 min at 37°C. Once fixed, coverslips were subjected to three 5-min washes in PBS at RT, after which cells were permeabilized in 0.1% Triton x-100 (Sigma-Aldrich) for 1 min at RT. After a single 5-min wash in PBS, samples were stained with 1 μm SYTOX Green for 5 min, followed by a 5-min wash in PBS. Once stained, specimens were mounted in fluoromount medium onto glass microscope slides (VWR International) and visualized using a LEICA DMI 6000 B microscope (Leica Microsystems, Milton Keynes, UK) at ×20 objective. The criterion used to define a NET was extracellular DNA in the form of a ‘line’ or ‘cloud’.

Hazeldine J., Harris P., Chapple I.L., Grant M., Greenwood H., Livesey A., Sapey E, & Lord J.M. (2014). Impaired neutrophil extracellular trap formation: a novel defect in the innate immune system of aged individuals. Aging Cell, 13(4), 690-698.

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Publication 2014

Atmosphere Biological Assay Cells Immunofluorescence Immunofluorescence Microscopy Microscopy Neutrophil paraform Reticular Formation SYTOX Green Triton X-100

Most recents protocols related to «SYTOX Green»

Membrane Permeability Assay of OCG Bactericidal Effect

Not available on PMC !

EXAMPLE 4

A membrane permeability assay using Sytox green dye was performed to examine whether the bactericidal effect is directly related to the disruption of membrane integrity. Fluorescence intensity of Sytox green increases when the membrane-impermeable dye intercalates into the intracellular nucleic acids upon diffusion through the damaged membranes. No fluorescence change was observed from Msm treated with OCG at 2×MIC for 1 h (FIG. 7). An additional assay commonly used for membrane damage was also conducted. Non-fluorescent hydrophobic N-phenyl-2-naphthylamine (NPN) becomes fluorescent upon interacting with damaged hydrophobic lipids in the membrane. Even after treating Msm with OCG at 4×MIC for 1 h, no fluorescence intensity increase was observed (FIG. 8). Both results indicated bactericidal effects of OCG may not be related to physical membrane damage.

US11857521B1. Anti-mycobacterial drugs (2024-01-02). None [US], None [US]. Inventors: Joong Ho Moon [US], Michelle Miranda [US].

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Patent 2024

Biological Assay Cell Membrane Permeability Diffusion Fluorescence Membrane Lipids Nucleic Acids Physical Examination Protoplasm SYTOX Green Tissue, Membrane

Neutrophil Extracellular Trap Formation Assay

Neutrophils of healthy donors were stimulated by PMA (25 nM). The impermeable DNA dye SYTOX Green was added immediately at a volume ratio of 1 : 100 per well and analysed using a cell real-time imaging system (Biotek, USA). Bright-field (phase) and fluorescence (GFP) channels were used to take pictures continuously. Each group was photographed for 4 h.

Han X., Zhang X., Song R., Li S., Zou S., Tan Q., Liu T., Luo S., Wu Z., Jie H, & Wang J. (2023). Necrostatin-1 Alleviates Diffuse Pulmonary Haemorrhage by Preventing the Release of NETs via Inhibiting NE/GSDMD Activation in Murine Lupus. Journal of Immunology Research, 2023, 4743975.

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Publication 2023

Cells Donors Fluorescence Neutrophil SYTOX Green

Neutrophil DNA Extrusion Monitoring

Neutrophils of healthy donors were stimulated using PMA (25 nM) for 2.5 h, and the impermeable DNA dye SYTOX Green (Thermo Fisher, USA, Cat No. S7020) was added to each well at a volume ratio of 1 : 100. Subsequently, fluorescence intensity was measured using a SpectraMax M5 multifunctional microplate reader (Biotek, USA) 10 min later. After PMA stimulation, the fluorescence intensity was recorded every 30 min for a total of 3.5 h.

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Publication 2023

Donors Fluorescence Neutrophil SYTOX Green

Neutrophil NET Formation Assay

Neutrophils of healthy donors were stimulated by PMA (25 nM) for 2.5 h, followed by the addition of the impermeable SYTOX Green DNA dye (Thermo Fisher, USA, Cat No. S7572) and the viable cell dye Hoechst 33342 (Beyotime, China, Cat No. C1025) at a volume ratio of 1 : 100 per well. NET production in each well was observed under an Olympus fluorescence microscope (Japan), and the randomly selected fields were photographed.

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Publication 2023

Cells Donors HOE 33342 Microscopy, Fluorescence Neutrophil SYTOX Green

Yeast Cell Fixation and DNA Staining Protocol

500μL samples were taken from yeast cultures (∼OD600 of 0.6). The cells were centrifuged at 3.000g for 2 min at RT. The supernatant was discarded and 1mL 70% Ethanol was added to the pellet. After thorough vortexing, the fixed cell suspensions can be stored at 4°C until further use. 500μL of the suspensions were transferred to a fresh tube and then centrifuged (3.000g, 2min RT). The supernatant was discarded and the pellet dissolved in 300μL 50mM sodium citrate and 0.1 mg/mL RNAse A. After 2h incubation at 50°C, proteinase K was added to a final concentration of 0.1 mg/mL, followed by another 2h incubation at 50°C. 30μL of this sample was then mixed with 170μL 50mM sodium citrate and 0.5μM Sytox Green (S7020, Thermo Fisher Scientific). Prior to FACS analysis, the samples were briefly sonicated (Bioruptor, 5 × 15sec) to detach cell clumps before proceeding with the analysis.

Weiβ M., Chanou A., Schauer T., Tvardovskiy A., Meiser S., König A.C., Schmidt T., Kruse E., Ummethum H., Trauner M., Werner M., Lalonde M., Hauck S.M., Scialdone A, & Hamperl S. (2023). Single-copy locus proteomics of early- and late-firing DNA replication origins identifies a role of Ask1/DASH complex in replication timing control. Cell Reports, 42(2), 112045.

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Publication 2023

Cells Endopeptidase K Endoribonucleases Ethanol Sodium Citrate SYTOX Green Yeast, Dried

Top products related to «SYTOX Green»

Sytox green by Thermo Fisher Scientific

Sourced in United States, Germany, United Kingdom, China, Switzerland, France, Canada, Netherlands, Spain, Italy, Austria

SYTOX Green is a nucleic acid stain that is membrane-impermeant, allowing it to selectively label dead cells with compromised plasma membranes. It exhibits a strong fluorescent signal upon binding to DNA.

Sytox green nucleic acid stain by Thermo Fisher Scientific

Sourced in United States, United Kingdom, Poland

SYTOX Green Nucleic Acid Stain is a fluorescent dye used for the detection and quantification of DNA and RNA in a variety of biological samples. It exhibits strong fluorescence upon binding to nucleic acids, making it a useful tool for flow cytometry, fluorescence microscopy, and other analytical techniques.

Phorbol myristate acetate (pma) by Merck Group

Sourced in United States, Germany, United Kingdom, France, Italy, China, Canada, Switzerland, Sao Tome and Principe, Macao, Poland, Japan, Australia, Belgium, Hungary, Netherlands, India, Denmark, Chile

The PMA is a versatile laboratory equipment designed for precision measurement and analysis. It functions as a sensitive pressure transducer, accurately measuring and monitoring pressure levels in various applications. The PMA provides reliable and consistent data for research and testing purposes.

Sytox green dye by Thermo Fisher Scientific

Sourced in United States, United Kingdom, Canada

SYTOX green dye is a nucleic acid stain used to detect cell death. It is membrane-impermeant and only stains cells with compromised membranes, making it useful for differentiating between live and dead cells.

Hoechst 33342 by Thermo Fisher Scientific

Sourced in United States, Germany, United Kingdom, Japan, China, France, Canada, Spain, Belgium, Italy, Australia, Austria, Denmark, Netherlands, Switzerland, Ireland, New Zealand, Portugal, Brazil, Argentina, Singapore, Poland, Ukraine, Macao, Thailand, Finland, Lithuania, Sweden

Hoechst 33342 is a fluorescent dye that binds to DNA. It is commonly used in various applications, such as cell staining and flow cytometry, to identify and analyze cell populations.

Sytox green dead cell stain by Thermo Fisher Scientific

Sourced in United States

SYTOX Green Dead Cell Stain is a fluorescent dye used for the detection of dead cells in a sample. It is a membrane-impermeant dye that binds to nucleic acids, providing a bright green fluorescence signal upon binding to DNA or RNA in cells with compromised membranes.

Triton x 100 by Merck Group

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Triton X-100 is a non-ionic surfactant commonly used in various laboratory applications. It functions as a detergent and solubilizing agent, facilitating the solubilization and extraction of proteins and other biomolecules from biological samples.

Rnase a by Merck Group

Sourced in United States, Germany, United Kingdom, France, Italy, China, Canada, Macao, Japan, Sao Tome and Principe, Spain, Switzerland, Belgium, India, Czechia, Australia, Israel

RNase A is a ribonuclease enzyme used in molecular biology laboratories. It functions by catalyzing the hydrolysis of single-stranded RNA, cleaving the phosphodiester bonds between nucleotides.

Incucyte zoom by Sartorius

Sourced in United States, United Kingdom, Germany, Japan

The IncuCyte ZOOM is a live-cell analysis system that enables real-time monitoring and quantification of cellular processes. It provides automated, non-invasive, and time-lapse imaging capabilities to observe and measure cellular responses within a controlled environment.

Facscalibur by BD

Sourced in United States, Germany, United Kingdom, China, Canada, Japan, Italy, France, Belgium, Switzerland, Singapore, Uruguay, Australia, Spain, Poland, India, Austria, Denmark, Netherlands, Jersey, Finland, Sweden

The FACSCalibur is a flow cytometry system designed for multi-parameter analysis of cells and other particles. It features a blue (488 nm) and a red (635 nm) laser for excitation of fluorescent dyes. The instrument is capable of detecting forward scatter, side scatter, and up to four fluorescent parameters simultaneously.

What is SYTOX Green and how is it used in biological research?

SYTOX Green is a fluorescent dye commonly used in various biological applications, particularly for detecting cell death and nucleic acid staining. It is a cell-impermeable dye, meaning it only enters cells with compromised membranes, making it a useful tool for identifying dead or dying cells. SYTOX Green binds to DNA, resulting in a bright green fluorescent signal that can be detected using fluorescence microscopy or flow cytometry. Researchers can leverage SYTOX Green to optimize their experimental protocols, enhance reproducibility, and gain accurate insights into their samples.

What are some common challenges or considerations when using SYTOX Green?

One key consideration when using SYTOX Green is ensuring that the cell membranes are sufficiently compromised to allow the dye to enter and bind to the DNA. This can be affected by factors such as cell type, experimental conditions, and the stage of cell death. Additionally, SYTOX Green can sometimes exhibit non-specific binding or background fluorescence, which can impact the accuracy of the results. Proper controls and optimization of the experimental protocol are crucial to mitigate these challenges and obtain reliable data.

Are there different variations or types of SYTOX Green available?

Yes, there are several variations of SYTOX Green available, each with slightly different characteristics and applications. For example, SYTOX Green nucleic acid stain is commonly used for detecting dead cells, while SYTOX Green dead cell stain is specifically designed for apoptosis and cell viability assays. Additionally, there are SYTOX Green variants with different excitation and emission wavelengths, allowing researchers to choose the dye that best fits their experimental setup and imaging equipment.

What are some specific applications of SYTOX Green in biological research?

SYTOX Green is widely used in a variety of biological applications, including: 1. Cell viability and cytotoxicity assays: SYTOX Green can be used to identify dead or dying cells, providing insights into the cytotoxic effects of drugs, chemicals, or environmental factors. 2. Apoptosis and programmed cell death studies: The dye can be used to detect and quantify apoptotic cells, enabling researchers to study the mechanisms and pathways involved in cell death. 3. Nucleic acid staining: SYTOX Green can be used to stain and visualize DNA or RNA, making it a useful tool for techniques like flow cytometry, fluorescence microscopy, and gel electrophoresis.

How can PubCompare.ai help optimize the use of SYTOX Green in research?

PubCompare.ai, an AI-driven platform, can help scientists easily locate relevant SYTOX Green protocols from literature, preprints, and patents, and use advanced comparisons to identify the best products and methods for their specific research needs. This can help improve the accuracy and reliability of SYTOX Green-based experiments and lead to more robust, reproducible results. The platform allows you to screen protocol literature more efficiently, leveraging AI to pinpoint critical insights that can help researchers identify the most effective SYTOX Green protocols for their research goals. The AI-driven analysis can highlight key differences in protocol effectiveness, enabling you to choose the best option for reproducibility and accuracy.

More about "SYTOX Green"

SYTOX Green is a versatile fluorescent dye widely used in various biological applications, particularly for detecting cell death and nucleic acid staining.
As a cell-impermeable dye, SYTOX Green only enters cells with compromised membranes, making it a valuable tool for identifying dead or dying cells.
The dye binds to DNA, resulting in a bright green fluorescent signal that can be detected using fluorescence microscopy or flow cytometry.
Researchers can leverage SYTOX Green to optimize their experimental protocols, enhance reproducibility, and gain accurate insights into their samples.
SYTOX Green is often compared to other nucleic acid stains like Hoechst 33342 and PMA (Propidium Monoazide), each with its own unique properties and applications.
To streamline the process of finding and optimizing SYTOX Green protocols, scientists can utilize AI-driven platforms like PubCompare.ai.
This innovative tool allows users to easily locate relevant SYTOX Green protocols from literature, preprints, and patents, and use advanced comparisons to identify the best products and methods for their specific research needs.
This can help improve the accuracy and reliability of SYTOX Green-based experiments, leading to more robust and reproducible results.
By leveraging the power of SYTOX Green and tools like PubCompare.ai, researchers can enhance their experimental workflows, maximize the impact of their findings, and drive scientific progress in their fields.
Whether you're studying cell death, nucleic acid dynamics, or other biological processes, SYTOX Green and PubCompare.ai can be invaluable resources in your research arsenal.