| Size | Price | Stock | Qty |
|---|---|---|---|
| 500mg |
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| 1g |
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| Other Sizes |
| Targets |
Methyl Green targets DNA by non-intercalative binding to the major groove of double-stranded DNA. The dye exhibits high affinity for DNA, with stable fluorescence emission upon binding. Its mechanism of action involves electrostatic interactions with the negatively charged phosphate backbone of DNA as well as sequence-specific binding to the major groove. The compound's fluorescence is enhanced upon DNA binding, allowing for sensitive detection of nucleic acids. Methyl Green is used as a DNA stain for cells and electrophoretic gels. In histological applications, it is used in Unna-Pappenheim staining coupled with Pyronin Y to visualize nucleic acids. The dye's resistance to photobleaching makes it particularly useful for fluorescence microscopy and flow cytometry applications.
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| ln Vitro |
Methyl Green exhibits potent DNA staining activity in vitro, with fluorescence excitation peaks at 633 nm and emission peaks at 677 nm. The dye demonstrates high affinity for DNA with minimal non-specific binding. In electrophoretic gels, Methyl Green can be used for direct visualization of DNA bands. The compound facilitates direct measurement of cell viability via microscopy and flow cytometry. It serves as a suitable counterstain for chloroacetate esterase, non-specific esterase, alkaline phosphatase, peroxidase, naphthol AS acetate esterase, and acid phosphatase. Methyl Green can be used for fluorescent labeling of nuclei in embryonic tissues or cells. The dye's stable fluorescence emission and resistance to photobleaching make it ideal for quantitative fluorescence-based assays.
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| ln Vivo |
Methyl Green is not used as a therapeutic agent and has no established in vivo pharmacological activity. It is primarily a research reagent used for staining and fluorescence applications. The compound is a histological dye used to visualize nucleic acids in tissue sections. It can be utilized for chromosome staining through the C-banding technique. Methyl Green has been used for fluorescent labeling of nuclei in embryonic tissues or cells. The dye's fluorescence properties make it suitable for flow cytometry applications in cell viability studies. As a research tool, Methyl Green is not administered to animals for therapeutic purposes but rather used ex vivo for staining and analytical applications.
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| Enzyme Assay |
In vitro experiments with Methyl Green typically involve preparing staining solutions at concentrations ranging from 0.1% to 1% w/v in appropriate buffers. For DNA staining in gel electrophoresis, the dye is added directly to the gel or used as a post-stain. For cell staining, cells are fixed and incubated with Methyl Green solution for 10-30 minutes, followed by washing steps. Fluorescence is measured using microscopy with excitation at 633 nm and emission at 677 nm. For flow cytometry, stained cells are analyzed using appropriate laser and filter settings. The dye is soluble in water and methanol. Staining specificity can be confirmed by DNase treatment, which eliminates Methyl Green fluorescence. Protection from light is essential during staining and imaging due to the dye's photosensitivity.
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| Cell Assay |
In vitro cell-based assays using Methyl Green involve staining of cultured cells or tissue sections. Cells are typically fixed with paraformaldehyde or methanol before staining. Methyl Green solution (0.1% w/v) is applied to the fixed cells and incubated for 10-30 minutes at room temperature. After washing with buffer or water, stained cells are visualized by fluorescence microscopy with excitation at 633 nm and emission at 677 nm. For flow cytometry analysis, stained cells are trypsinized, resuspended in buffer, and analyzed using appropriate laser settings. The dye can be used for viability assessment as dead cells may exhibit altered staining patterns. For nuclear labeling in embryonic tissues, the dye is applied to tissue sections following standard histological protocols. The staining can be combined with other dyes for multiplex analysis.
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| Animal Protocol |
Methyl Green is not typically used for in vivo animal experiments as a therapeutic agent. However, the compound has been used in ex vivo staining of tissues from animal models. For histological analysis, tissues are harvested from animals, fixed, sectioned, and stained with Methyl Green for nuclear visualization. The dye can be used for chromosome staining by C-banding technique in cytogenetic studies. For fluorescence imaging, tissues may be stained with Methyl Green and examined under a fluorescence microscope. The dye is not administered to live animals for therapeutic purposes but rather used as a research tool for tissue analysis. Standard protocols for tissue fixation, sectioning, and staining are followed, with Methyl Green used at concentrations of 0.1-1% w/v in appropriate buffers.
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| ADME/Pharmacokinetics |
Methyl Green does not have established pharmacokinetic properties as it is not used as a therapeutic agent. The compound is a histological dye and fluorescent stain used for research applications. For in vitro use, Methyl Green is soluble in water and methanol, with solutions typically prepared at 0.1% w/v. The dye exhibits fluorescence excitation at 633 nm and emission at 677 nm. For storage, Methyl Green powder should be kept at -20°C for up to 3 years, and solutions should be stored at -80°C for up to 1 year. Protection from light is imperative to maintain the dye's fluorescence properties. The compound is stable under recommended storage conditions and should be protected from direct sunlight.
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| Toxicity/Toxicokinetics |
The toxicity of Methyl Green is primarily associated with its use as a dye rather than as a therapeutic agent. The compound is intended for research use only and is not for human use. As a histological dye, Methyl Green should be handled with appropriate laboratory safety precautions, including the use of gloves and eye protection. The compound may cause irritation to skin, eyes, and respiratory tract upon contact. Inhalation of dust should be avoided. The dye is not classified as a highly toxic substance but should be handled in a well-ventilated area. Standard safety data sheets recommend washing affected areas with soap and water in case of skin contact and seeking medical attention if irritation persists. Chronic toxicity data are limited as the compound is not used in therapeutic applications.
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| References | |
| Additional Infomation |
Ethyl green is an imine salt composed of [4-([4-(dimethylamino)phenyl]{4-[ethyl(dimethyl)azono]phenyl}methylene)cyclohexyl-2,5-diene-1-methylene](dimethyl)ammonium, bromide ions, and chloride ions in a 1:1:1 ratio. It is a histological dye used to visualize nucleic acids by Unna-Pappenheim staining coupled with Pyronin Y. It is both a fluorescent dye and a histological dye. It is an organochloride salt, quaternary ammonium salt, imine salt, and organobromine salt. It contains ethyl green (1+).
See also: Methyl green (note moved to). Methyl Green (CAS 14855-76-6) is a basic dye useful for chromatin staining. It is also known by the UNII code QFQ95SNB21 and C.I. 42590. The term "methyl green" also applies to the trimethyl analog. The dye is used in textile dyeing and printing in addition to biological staining. Methyl Green is a non-intercalating fluorescent dye that selectively binds to the major groove of DNA. It exhibits high affinity for DNA, resistance to photobleaching, and stable fluorescence emission. The dye is used in Unna-Pappenheim staining coupled with Pyronin Y to visualize nucleic acids. Methyl Green is both a fluorescent dye and a histological dye. It is available as a 0.1% w/v aqueous solution and as a powder. The compound should be protected from light during storage. |
| Molecular Formula |
C27H35N3BRCL
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|---|---|
| Molecular Weight |
516.9439
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| Exact Mass |
457.205
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| CAS # |
14855-76-6
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| PubChem CID |
84671
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| Appearance |
Green to dark green solid powder
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
3
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
32
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| Complexity |
645
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CC[N+](C)(C)C1=CC=C(C=C1)C(=C2C=CC(=[N+](C)C)C=C2)C3=CC=C(C=C3)N(C)C.[Cl-].[Br-]
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| InChi Key |
IDAQSADEMXDTKN-UHFFFAOYSA-L
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| InChi Code |
InChI=1S/C27H35N3.BrH.ClH/c1-8-30(6,7)26-19-13-23(14-20-26)27(21-9-15-24(16-10-21)28(2)3)22-11-17-25(18-12-22)29(4)5;;/h9-20H,8H2,1-7H3;2*1H/q+2;;/p-2
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| Chemical Name |
[4-[[4-(dimethylamino)phenyl]-(4-dimethylazaniumylidenecyclohexa-2,5-dien-1-ylidene)methyl]phenyl]-ethyl-dimethylazanium;bromide;chloride
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : ~125 mg/mL (~241.81 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (4.02 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.08 mg/mL (4.02 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.9345 mL | 9.6723 mL | 19.3446 mL | |
| 5 mM | 0.3869 mL | 1.9345 mL | 3.8689 mL | |
| 10 mM | 0.1934 mL | 0.9672 mL | 1.9345 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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