2. Cell Cycle/DNA Damage
  3. DNA Stain
  4. Methyl Green

Methyl Green is a non-intercalating fluorescent labeling agent that selectively binds to the major groove of DNA. Methyl Green electrostatically interacts with the major groove of DNA through positively charged groups, exhibiting key activities such as high affinity, resistance to photobleaching, and stable fluorescence emission. Methyl Green can be directly measured by microscopy and flow cytometry, with peaks at 633 and 677 nm. Methyl Green can be used for fluorescent labeling of the nuclei of embryonic tissues or cells, or DNA staining and cell activity detection in gel electrophoresis[1][2][3].

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Methyl Green Chemical Structure

Methyl Green Chemical Structure

CAS No. : 14855-76-6

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Based on 1 publication(s) in Google Scholar

Methyl Green Related Antibodies

Top Publications Citing Use of Products

1 Publications Citing Use of MCE Methyl Green

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  • Purity & Documentation

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Description

Methyl Green is a non-intercalating fluorescent labeling agent that selectively binds to the major groove of DNA. Methyl Green electrostatically interacts with the major groove of DNA through positively charged groups, exhibiting key activities such as high affinity, resistance to photobleaching, and stable fluorescence emission. Methyl Green can be directly measured by microscopy and flow cytometry, with peaks at 633 and 677 nm. Methyl Green can be used for fluorescent labeling of the nuclei of embryonic tissues or cells, or DNA staining and cell activity detection in gel electrophoresis[1][2][3].

In Vitro

I. Zebrafish/chicken embryo whole embryo fluorescence staining reference[1][2]
Steps:
1. Sample fixation: 48 hpf zebrafish embryos or target stage chicken embryos were fixed with 4% paraformaldehyde (HY-Y0333) (prepared with PBS) at 4°C overnight. The fixation time was adjusted according to the thickness of the embryo.
2. Permeabilization: The embryos were washed 3 times with PBS (PBS-T) (HY-K1022) containing 1% Triton X-100 for 5 minutes each time to destroy the egg membrane/tissue barrier.
3. Methyl green staining: dilute 2% methyl green stock solution to 1:5,000-1:10,000 with PBS-T (final concentration of about 2-4 μg/mL), incubate embryos at 4°C for at least 6 hours (overnight for thick embryos), with gentle shaking.
4. Washing and removal of impurities: wash 3 times with PBS, 30 minutes each time, to remove unbound dye.
5. Mounting and imaging: mount the slides with 75% glycerol + 0.1 M Tris·HCl (pH 8.0), use laser confocal microscopy, excitation wavelength 633 nm, emission wavelength 650-750 nm (peak 677 nm).

II. Agarose gel DNA staining[2]
Steps:
1. Gel preparation: Add 2% Methyl Green (HY-D0163) stock solution to 1% agarose (prepared with TAE buffer) at a ratio of 1:5,000-1:10,000, and cool after gel preparation.
2. Sample loading: Load DNA/RNA ladder (such as 100 bp DNA ladder), conventional electrophoresis (voltage/time adjusted according to the experiment).
3. Fluorescence detection: Use 635 nm red light excitation, 705 nm bandpass filter, exposure for 30-60 seconds, or observe with UV transilluminator (only DNA color development).

III. Flow cytometry to detect cell viability[2][3]
Steps:
1. Cell treatment: Cultured Raji cells (or PBMCs) were treated with drugs (e.g. 50-100 μg/mL Fludarabine (HY-B0069)) for 24 hours, and then the cell suspension (5×105 cells) was collected.
2. Staining: Add 4 μg/mL Methyl Green (HY-D0163) + 10 μg/mL Propidium Iodide (HY-D0815) and incubate at room temperature in the dark for 15 minutes.
3. Flow cytometry: Use a CyAn flow cytometer, set the excitation wavelength to 633 nm (MG)/488 nm (PI), and the emission wavelength to 677 nm (MG)/609 nm (PI), and analyze the ratio of live and dead cells.

IV. Fluorescence spectroscopy (DNA binding analysis)[2]
Chinese steps:
1. Solution preparation: Prepare a 100 μg/mL calf thymus DNA (HY-109517) solution in PBS and add methyl green to a final concentration of 10 μg/mL (or 125 ng/mL free state).
2. Spectral detection: Use a fluorescence spectrophotometer, excite at 633 nm, scan the emission spectrum from 645-800 nm, and record the peak at 677 nm (DNA binding state).

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Methyl Green Related Antibodies
Molecular Weight

516.94

Formula

C27H35BrClN3
CAS No.
Appearance

Solid

Color

Green to dark green

SMILES

C[N+](C)(CC)C1=CC=C(/C(C2=CC=C(N(C)C)C=C2)=C3C=C/C(C=C\3)=[N+](C)\C)C=C1.[Br-].[Cl-]
Shipping

Room temperature in continental US; may vary elsewhere.
Storage

4°C, sealed storage, away from moisture and light

*In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)

Solvent & Solubility
In Vitro: 

DMSO : 125 mg/mL (241.81 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

Preparing
Stock Solutions
Concentration Solvent Mass 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
View the Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

  • Molarity Calculator

  • Dilution Calculator

Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

Mass
=
Concentration
×
Volume
×
Molecular Weight *

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

This equation is commonly abbreviated as: C1V1 = C2V2

Concentration (start)

C1

×
Volume (start)

V1

=
Concentration (final)

C2

×
Volume (final)

V2

In Vivo:

Select the appropriate dissolution method based on your experimental animal and administration route.

For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

  • Protocol 1

    Add each solvent one by one:  10% DMSO    40% PEG300    5% Tween-80    45% Saline

    Solubility: ≥ 2.08 mg/mL (4.02 mM); Clear solution

    This protocol yields a clear solution of ≥ 2.08 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (20.8 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.

    Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
  • Protocol 2

    Add each solvent one by one:  10% DMSO    90% (20% SBE-β-CD in Saline)

    Solubility: ≥ 2.08 mg/mL (4.02 mM); Clear solution

    This protocol yields a clear solution of ≥ 2.08 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (20.8 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.

    Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.
In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:

Dosage

mg/kg

Animal weight
(per animal)

g

Dosing volume
(per animal)

μL

Number of animals

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
%
DMSO +
% 
+
%
Tween-80 +
%
Saline
Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).
Calculation results:
Working solution concentration: mg/mL
Method for preparing stock solution: mg drug dissolved in μL  DMSO (Stock solution concentration: mg/mL).

*In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)

The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only. If necessary, please contact MedChemExpress (MCE).
Method for preparing in vivo working solution for animal experiments: Take μL DMSO stock solution, add μL . μL , mix evenly, next add μL Tween 80, mix evenly, then add μL Saline.
 If the continuous dosing period exceeds half a month, please choose this protocol carefully.
Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
Purity & Documentation

Purity: 82.50%

SDS (923 KB)

COA (250 KB)

Handling Instructions (2659 KB)
References

Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
DMSO 1 mM 1.9345 mL 9.6723 mL 19.3446 mL 48.3615 mL
5 mM 0.3869 mL 1.9345 mL 3.8689 mL 9.6723 mL
10 mM 0.1934 mL 0.9672 mL 1.9345 mL 4.8362 mL
15 mM 0.1290 mL 0.6448 mL 1.2896 mL 3.2241 mL
20 mM 0.0967 mL 0.4836 mL 0.9672 mL 2.4181 mL
25 mM 0.0774 mL 0.3869 mL 0.7738 mL 1.9345 mL
30 mM 0.0645 mL 0.3224 mL 0.6448 mL 1.6121 mL
40 mM 0.0484 mL 0.2418 mL 0.4836 mL 1.2090 mL
50 mM 0.0387 mL 0.1934 mL 0.3869 mL 0.9672 mL
60 mM 0.0322 mL 0.1612 mL 0.3224 mL 0.8060 mL
80 mM 0.0242 mL 0.1209 mL 0.2418 mL 0.6045 mL
100 mM 0.0193 mL 0.0967 mL 0.1934 mL 0.4836 mL
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Methyl Green Related Classifications

Help & FAQs
  • Do most proteins show cross-species activity?

    Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

Keywords:

Methyl Green14855-76-6DNA Stainfluorescent dyeDNAcell viabilitymicroscopyflow cytometryInhibitorinhibitorinhibit

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