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Methylene Blue Powder, Cat. No. ASD-616
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Methylene Blue Powder
Laboratory Staining Dye & Redox Indicator — CAS 61-73-4
Cat. No. ASD-616 Methylthioninium Chloride C₁₆H₁₈ClN₃S MW 319.85 g/mol CI 52015 | Thiazine Dye 🇦🇺 Made in Melbourne ⚡ Same-Week Dispatch
Overview & Biochemistry
Methylene Blue (Methylthioninium Chloride) is a cationic thiazine dye with one of the broadest application profiles of any laboratory reagent. Its deep blue coloration in oxidised form, excellent water solubility, and well-characterised reversible redox behaviour make it indispensable across microbiology, histology, molecular biology, biochemistry, and electrochemical research.
Supplied as a dark green to bronze crystalline powder with metallic lustre — this distinctive appearance is characteristic of high-purity Methylene Blue and results from the strong absorption of the oxidised form in solution (~664 nm).
Mode of Action
Cationic Staining → Nucleic Acid & Polyanion Binding | Reversible Redox: MB⁺ ⇄ MBH (Leucomethylene Blue)
As a cationic dye, Methylene Blue binds electrostatically to negatively charged cellular structures — particularly nucleic acids (RNA > DNA), acidic mucopolysaccharides, and bacterial cell walls — producing intense blue staining. In redox applications, MB undergoes a fully reversible one-electron/two-proton reduction: the deep blue oxidised form (MB⁺) is reduced to colourless leucomethylene blue (MBH) by cellular reductants or applied reducing agents. This reversible colour change (blue ↔ colourless) is the basis of its use as a redox indicator, electron mediator in biosensors, and viability dye in certain biological systems.
As a cationic dye, Methylene Blue binds electrostatically to negatively charged cellular structures — particularly nucleic acids (RNA > DNA), acidic mucopolysaccharides, and bacterial cell walls — producing intense blue staining. In redox applications, MB undergoes a fully reversible one-electron/two-proton reduction: the deep blue oxidised form (MB⁺) is reduced to colourless leucomethylene blue (MBH) by cellular reductants or applied reducing agents. This reversible colour change (blue ↔ colourless) is the basis of its use as a redox indicator, electron mediator in biosensors, and viability dye in certain biological systems.
Applications
🧪
Bacterial Staining
Simple stain for cell morphology
🔬
RNA Visualisation
Northern blot staining; RNA gels
⚡
Redox Indicator
Biochemical & electrochemical assays
📊
Biosensor Research
Electron mediator in electrodes
🏭
Wastewater Research
Dye degradation model compound
🏫
Education
Redox demos; cell staining labs
Simple Bacterial Staining Protocol
| 1 | Prepare smear: Spread a small amount of culture on a clean glass slide. Air dry completely, then heat-fix by passing through a flame 2–3 times. |
| 2 | Prepare staining solution: Dissolve Methylene Blue (ASD-616) in distilled water to make a 0.3% w/v solution (Loeffler's Methylene Blue: add 0.01% KOH to intensify). Filter through Whatman No. 1 before use. |
| 3 | Stain: Flood slide with Methylene Blue solution. Leave for 1–3 minutes. |
| 4 | Rinse: Gently rinse with distilled water until rinse runs clear. Blot dry — do not rub. |
| 5 | Examine: View under oil immersion (×1000). Bacteria stain blue; background remains pale. Useful for cell morphology, size estimation, and metachromatic granule detection in Corynebacterium spp. |
RNA Gel Staining Protocol (Northern Blot / Agarose Gel)
| 1 | After transfer, incubate membrane in 0.04% Methylene Blue in 0.5 M sodium acetate (pH 5.2) for 5–10 minutes with gentle agitation. |
| 2 | Destain in distilled water with several changes until background clears (typically 10–30 min). RNA bands appear as deep blue bands on a pale background. |
| 3 | Photograph membrane. Staining is reversible — membranes can be destained fully and reprobed with hybridisation probes after MB staining. |
⚠️ MB RNA staining is a rapid loading control method but is less sensitive than EtBr or SYBR dyes for gel visualisation. Use 0.02–0.04% concentration range; higher concentrations increase background.
Cross-Reference / Equivalent Products
| Manufacturer | Product Name | Cat. No. | Notes |
|---|---|---|---|
| Sigma-Aldrich (Merck) | Methylene Blue | M9140 | Equivalent powder grade |
| Merck Millipore | Methylene Blue for microscopy (Certistain®) | 1.01290 | Equivalent powder grade |
| Thermo Fisher Scientific | Methylene Blue Solution | M291 | Ready-to-use solution; ASD-616 is powder |
| Oxoid (Thermo Fisher) | Methylene Blue (Loeffler's) | BR0099 | Ready-to-use staining reagent |
Cross-references are for convenience only. Catalogue numbers are trademarks of their respective owners.
Related Products
Chemical & Physical Properties
| Product Name | Methylene Blue Powder |
| Catalogue Number | ASD-616 |
| IUPAC Name | 3,7-bis(dimethylamino)phenothiazin-5-ium chloride (Methylthioninium Chloride) |
| Synonyms | Basic Blue 9; CI 52015; Tetramethylthionine Chloride; Methylthioninium Chloride |
| CAS Number | 61-73-4 |
| Colour Index | CI 52015 |
| Molecular Formula | C₁₆H₁₈ClN₃S |
| Molecular Weight | 319.85 g/mol |
| Dye Class | Thiazine dye (cationic) |
| Grade | Laboratory Grade |
| Appearance | Dark green to bronze crystalline powder with metallic lustre |
| Dye Content | ≥82% (anhydrous basis, CI 52015) |
| Solubility (water) | ~40 g/L at 25°C (highly soluble) |
| Solubility (ethanol) | Soluble (~10 g/L) |
| Solubility (chloroform) | Slightly soluble |
| λmax (oxidised form) | ~664 nm (aqueous solution) |
| Redox Potential (E°) | +0.011 V vs SHE (MB⁺/MBH couple, pH 7) |
| Melting Point | >180°C (with decomposition) |
Working Solution Concentrations by Application
| Application | Concentration | Solvent | Notes |
|---|---|---|---|
| Simple bacterial stain | 0.3% w/v | Distilled water | Loeffler's: add 0.01% KOH |
| RNA membrane staining | 0.02–0.04% w/v | 0.5 M NaOAc pH 5.2 | Northern blot loading control |
| Redox indicator solution | 0.01–0.1% w/v | Distilled water | Decolourises on reduction |
| Electrochemical mediator | 0.1–1.0 mM | Buffer (pH 7) | E° = +0.011 V vs SHE |
| Dye degradation model | 10–100 mg/L | Distilled water | Standard photocatalysis substrate |
Storage & Stability
| Storage Temperature | 15–25°C (ambient, away from heat) |
| Container | Tightly sealed, light-resistant container |
| Protect From | Moisture, direct light, and strong reducing agents |
| Shelf Life (powder) | 3 years from manufacture date when stored correctly |
| Working Solutions | Aqueous solutions stable 12 months in sealed dark bottle at RT |
Literature & References
| # | Reference |
|---|---|
| 1 | Wainwright M. Methylene blue derivatives — suitable photoantimicrobials for blood product disinfection? Int J Antimicrob Agents. 2000;16(4):381–394. [Broad review of MB chemistry and applications] |
| 2 | Sambrook J, Russell DW. Molecular Cloning: A Laboratory Manual. 3rd ed. Cold Spring Harbor Laboratory Press; 2001. [RNA staining protocol with methylene blue] |
| 3 | Loeffler F. Untersuchungen über die Bedeutung der Mikroorganismen für die Entstehung der Diphtherie. Mitt Kaiserl Gesundh. 1884;2:421–499. [Original Loeffler's methylene blue formulation] |
| 4 | Phenrat T, Saleh N, Sirk K, et al. Aggregation and sedimentation of aqueous nanoscale zerovalent iron dispersions. Environ Sci Technol. 2007;41(1):284–290. [MB as model dye in environmental remediation studies] |
| 5 | Ramirez AG, et al. Methylene blue as electrochemical mediator in biosensor design. Biosens Bioelectron. 2019;126:103–115. [Redox mediator applications] |
📄 Full SDS available: Download the complete 16-section GHS Safety Data Sheet (Australian WHS Regulations 2023 / GHS 7th Edition) — contact support@ausamics.com.au
Section 1 — Identification
| Product Name | Methylene Blue Powder |
| Catalogue No. | ASD-616 |
| Supplier | AuSaMicS Pty Ltd | ABN 56 676 640 467 |
| Address | 31 Longview CT, Thomastown VIC 3074, Australia |
| Emergency | Poisons Information Centre: 13 11 26 (24 hr, Australia) |
| Phone | +61 412 520 598 |
| support@ausamics.com.au |
Section 2 — Hazard Identification (GHS 7th Ed / WHS 2023)
| GHS Classification | Acute Tox. 4 (oral) — H302: Harmful if swallowed Eye Irrit. 2 — H319: Causes serious eye irritation Skin Sens. 1B — H317: May cause an allergic skin reaction Aquatic Chronic 3 — H412: Harmful to aquatic life with long-lasting effects |
| Signal Word | WARNING |
| GHS Pictograms | ⚠️ Exclamation mark | 🌿 Environmental hazard |
| Practical Note | Stains skin, mucous membranes, and clothing blue — use nitrile gloves and eye protection. Staining is persistent but not hazardous. |
Sections 4–8 — First Aid, Fire & PPE
| Inhalation | Remove to fresh air. Seek medical attention if symptoms persist. Avoid generating dust. |
| Skin Contact | Wash with soap and water ≥15 min. Blue skin staining is expected — fades within days. Seek advice if irritation or rash develops. |
| Eye Contact | Irrigate with large amounts of water ≥15 min. Seek medical attention immediately if irritation persists. |
| Ingestion | Do NOT induce vomiting. Rinse mouth. Call Poisons Centre 13 11 26. Seek medical attention. Note: urine may turn blue — this is expected and not dangerous at small amounts. |
| Extinguishing Media | Water spray, foam, dry powder, CO₂ |
| Respiratory Protection | P1/P2 particulate filter when handling bulk powder |
| Eye Protection | Safety glasses or splash goggles |
| Skin Protection | Nitrile gloves (≥0.11 mm), lab coat, closed footwear |
Sections 12–15 — Ecology, Disposal & Regulatory
| Aquatic Toxicity | Harmful to aquatic life with long-lasting effects (H412). Do not discharge to waterways. |
| Disposal | Dispose per local EPA regulations as chemical waste. Do not discharge to stormwater drains. |
| Transport | Not classified as dangerous goods — ADG Code, IMDG, IATA |
| Regulatory Framework | Australian WHS Regulations 2023 | GHS 7th Edition | Safe Work Australia COP 2021 |
| AICIS Status | Listed on Australian Inventory of Industrial Chemicals (AIIC) |
| Poisons Schedule | Not scheduled under SUSMP for laboratory/research use quantities |
Quality Specifications
| Parameter | Specification | Method |
|---|---|---|
| Appearance | Dark green to bronze crystalline powder with metallic lustre | Visual |
| Dye Content | ≥82% (anhydrous, CI 52015) | Dye content titration |
| λmax | 664 ± 4 nm | UV-Vis spectrophotometry |
| Solubility | Complete dissolution in distilled water at 10 mg/mL; deep blue clear solution | Visual inspection |
| pH (0.1% aq. solution) | 3.0–4.5 | pH meter |
| Heavy Metals | ≤20 ppm | ICP-OES |
| Batch COA | Available for every production lot | On request or download |
Manufacturing & Documentation
Manufactured by AuSaMicS Pty Ltd
31 Longview CT, Thomastown VIC 3074, Australia | ABN 56 676 640 467
✓ Batch-level quality control on every lot
✓ Full GHS-compliant documentation (COA, TDS, SDS)
✓ Australian stock — no import delays, no customs holdups
✓ Technical support direct from our formulation team
✓ Available in gram-scale to kilogram-scale quantities
31 Longview CT, Thomastown VIC 3074, Australia | ABN 56 676 640 467
✓ Batch-level quality control on every lot
✓ Full GHS-compliant documentation (COA, TDS, SDS)
✓ Australian stock — no import delays, no customs holdups
✓ Technical support direct from our formulation team
✓ Available in gram-scale to kilogram-scale quantities
Available Documentation
| 📄 Certificate of Analysis (COA) | Batch-specific | Available on request |
| 📋 Technical Data Sheet (TDS) | Full specifications and preparation notes |
| ⚠️ Safety Data Sheet (SDS) | GHS 7th Ed | WHS 2023 compliant | 16 sections |
⚠️ Intended Use: For laboratory research and educational use only. Not for human consumption, veterinary, agricultural, diagnostic, therapeutic, food, feed, cosmetic, or pharmaceutical use. International buyers must confirm import eligibility with their local regulatory authorities.