Methylene Blue (Synonyms: Basic Blue 9; CI-52015; Methylthioninium chloride)

Name: Methylene Blue
Brand: MedChemExpress
SKU: HY-14536
Rating: 5.0 (28 reviews)

Cat. No.: HY-14536 Purity: 95.05%: Data Sheet
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Methylene blue (Basic Blue 9) is a guanylyl cyclase (sGC), monoamine oxidase A (MAO-A) and NO synthase (NOS) inhibitor. Methylene blue is a vasopressor and is often used as a dye in several medical procedures. Methylene blue through the nitric oxide syntase/guanylate cyclase signalling pathway to reduce prepulse inhibition. Methylene blue is a REDOX cycling compound and able to cross the blood-brain barrier. Methylene blue is a Tau aggregation inhibitor. Methylene Blue is a photosensitizer and redox agent. Methylene blue reduces cerebral edema, attenuated microglial activation and reduced neuroinflammation.

For research use only. We do not sell to patients.

CAS No. : 61-73-4

Size	Stock	Quantity
Solid + Solvent (Highly Recommended)
10 mM * 1 mL in Water ready for reconstitution	In-stock	Estimated Time of Arrival: December 31
Solution
10 mM * 1 mL in Water	In-stock	Estimated Time of Arrival: December 31
Solid
100 mg	In-stock	Estimated Time of Arrival: December 31
500 mg	In-stock	Estimated Time of Arrival: December 31
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Based on 28 publication(s) in Google Scholar

Other Forms of Methylene Blue:

28 Publications Citing Use of MCE Methylene Blue

Gel Electrophoresis

Cell Migration/Invasion Assay

: Methylene Blue purchased from MedChemExpress. Usage Cited in: Nat Commun. 2024 Nov 27;15(1):10303. [Abstract]; Northern blot analysis of pre-rRNA intermediates using 5'ETS-1, 5'ETS-2, 5'ETS and ITS1-29 probes in DDX10-AID mESCs with IAA treatment (0 h, 6 h, 24 h, and 48 h) or treated with IAA for 48 h followed by 48 h of washing. MB represents methylene blue staining. Experiments were repeated three times independently with similar results. Membranes were stained with Methylene Blue stain solution (0.03% Methylene Blue in 0.3 M sodium acetate (pH 5.0-5.5)), and then RNA was crosslinked onto the membrane using UV light.

: Methylene Blue purchased from MedChemExpress. Usage Cited in: Nat Commun. 2024 Nov 27;15(1):10303. [Abstract]; Northern blot analysis of pre-rRNA intermediate levels in DDX10-AID (+ OsTir1) mESCs overexpressing DDX10FL, DDX10ΔHBD, DDX10ΔHCD or DDX10ΔHD at 0 h and 48 h after IAA treatment. MB represents Methylene Blue staining. Experiments were repeated three times independently with similar results. Source data are provided as a Source Data file. Membranes were stained with Methylene Blue stain solution (0.03% Methylene Blue in 0.3 M sodium acetate (pH 5.0-5.5)), and then RNA was crosslinked onto the membrane using UV light.

: Methylene Blue purchased from MedChemExpress. Usage Cited in: Nat Commun. 2024 Nov 27;15(1):10303. [Abstract]; Northern blot analysis of pre-rRNA intermediates in DDX10-AID (+ OsTir1) mESCs overexpressing NUP98-DDX10 or DDX10, treated with IAA at different time points (0 h, 24 h, and 48 h). MB represents Methylene Blue staining. Experiments were repeated two times independently with similar results. Source data are provided as a Source Data file. Membranes were stained with Methylene Blue stain solution (0.03% Methylene Blue in 0.3 M sodium acetate (pH 5.0-5.5)), and then RNA was crosslinked onto the membrane using UV light.

: Methylene Blue purchased from MedChemExpress. Usage Cited in: Theranostics. 2021 Oct 17;11(20):9884-9903. [Abstract]; Migration analysis of THP-1, co-cultured with cancer cells that were pre-treated with Methylene Blue.

: Methylene Blue purchased from MedChemExpress. Usage Cited in: Theranostics. 2021 Oct 17;11(20):9884-9903. [Abstract]; Western blotting analysis of PD-L1 expression in THP-1 which was co-cultured with cancer cells pre-treated with Methylene Blue.

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Biological Activity
Purity & Documentation
References
Customer Review

Description

IC₅₀ & Target

Guanylyl cyclase (sGC)^[1].
Monoamine oxidase A (MAO-A)^[1].
NO synthase (NOS)^[1]

Cellular Effect

Cell Line	Type	Value	Description	References
B16-F10	IC₅₀	5.48 μM Compound: MB	Antiproliferative activity against mouse B16-F10 cells assessed as cell viability in light irradiation and measured for 48 hrs by MTT assay Antiproliferative activity against mouse B16-F10 cells assessed as cell viability in light irradiation and measured for 48 hrs by MTT assay	[PMID: 34871840]
B16-F10	IC₅₀	7.02 μM Compound: MB	Antiproliferative activity against mouse B16-F10 cells assessed as cell viability in dark incubated for 48 hrs by MTT assay Antiproliferative activity against mouse B16-F10 cells assessed as cell viability in dark incubated for 48 hrs by MTT assay	[PMID: 34871840]
FM3A	EC₅₀	1.1 x 10^-6 M Compound: Methylene blue	Cytotoxicity against mouse mammary tumor FM3A cells representing a model of host Cytotoxicity against mouse mammary tumor FM3A cells representing a model of host	[PMID: 11855978]
Fibroblast	EC₅₀	43 nM Compound: MB	Inhibition of erastin-induced cell death in FRDA patient-derived fibroblast assessed as depletion of cellular ATP incubated for 12 hrs followed by erastin stimulation and measured after by luciferase-linked ATPase enzymatic assay Inhibition of erastin-induced cell death in FRDA patient-derived fibroblast assessed as depletion of cellular ATP incubated for 12 hrs followed by erastin stimulation and measured after by luciferase-linked ATPase enzymatic assay	[PMID: 33214825]
HT-29	IC₅₀	9.2 μM Compound: MB	Cytotoxicity against human HT-29 cells assessed as reduction in cell viability preincubated for 1 hr followed by light irradiation at 3.2 J/cm2 for 30 sec and measured after 24 hrs by MTT assay Cytotoxicity against human HT-29 cells assessed as reduction in cell viability preincubated for 1 hr followed by light irradiation at 3.2 J/cm2 for 30 sec and measured after 24 hrs by MTT assay	[PMID: 32267685]
HT-29	IC₅₀	> 10 μM Compound: MB	Cytotoxicity against human HT-29 cells assessed as reduction in cell viability incubated for 24 hrs in dark by MTT assay Cytotoxicity against human HT-29 cells assessed as reduction in cell viability incubated for 24 hrs in dark by MTT assay	[PMID: 32267685]
HepG2	IC₅₀	6 μM Compound: MB	Anticancer activity against human HepG2 cells Anticancer activity against human HepG2 cells	[PMID: 33460833]
HepG2	IC₅₀	< 1 μM Compound: Methylene blue	Antimalarial activity against sporozoite stage of Plasmodium yoelii assessed as invasion of human HepG2 cells expressing CD81 incubated for 2 hrs prior to inoculation measured after 1 hr by immunofluorescence assay in presence of penicillin/streptomycin Antimalarial activity against sporozoite stage of Plasmodium yoelii assessed as invasion of human HepG2 cells expressing CD81 incubated for 2 hrs prior to inoculation measured after 1 hr by immunofluorescence assay in presence of penicillin/streptomycin	[PMID: 23927658]
Lymphocyte	EC₅₀	88 nM Compound: MB	Anti-ferroptotic activity in FRDA patient-derived Lymphocyte assessed as reduction in RSL3-induced lipid peroxidation incubated for overnight followed by RSL3 stimulation and measured after 90 mins by FACS analysis Anti-ferroptotic activity in FRDA patient-derived Lymphocyte assessed as reduction in RSL3-induced lipid peroxidation incubated for overnight followed by RSL3 stimulation and measured after 90 mins by FACS analysis	[PMID: 33214825]
MCF7	IC₅₀	10.58 μM Compound: MB	Antiproliferative activity against human MCF7 cells assessed as cell viability in light irradiation and measured after 48 hrs by MTT assay Antiproliferative activity against human MCF7 cells assessed as cell viability in light irradiation and measured after 48 hrs by MTT assay	[PMID: 34871840]
MCF7	IC₅₀	14.64 μM Compound: MB	Antiproliferative activity against human MCF7 cells assessed as cell viability in dark incubated for 48 hrs by MTT assay Antiproliferative activity against human MCF7 cells assessed as cell viability in dark incubated for 48 hrs by MTT assay	[PMID: 34871840]

In Vitro

Methylene blue (Basic Blue 9) (4.5 μM; BV2 microglia) alters the immune profile of LPS-activated BV2 microglia and decreases the level of CD14, IL-1β, TNF-α, and CCL2 mRNA^[3].
1. Sample treatment:
1.1 For paraffin sections: dewax and rehydrate as usual.
1.2 For frozen sections: distilled water for 2 min.
1.3 For cultured cells: Fix with 4% paraformaldehyde. Wash with distilled water for 2 min. Replace with fresh distilled water and wash for another 2 min.
2. Methylene blue staining
2.1 Stain with 0.1% Methylene blue for 2 min.
2.2 Wash thoroughly with distilled water, observe and take photos.
2.3 Staining results:
Tissue or cells: blue

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

In Vivo

Methylene blue (Basic Blue 9) (50 and 100 mg/kg; i.p.; once, for 25 min; male NMRI mice) reduces absent prepulse inhibition^[1].
Methylene blue (Basic Blue 9) (20 and 40 mg/kg; p.o.; daily, for 6 months; CaMKIIα-tTA transactivator mice) preserves cognition in mice expressing full-length pro-aggregant human Tau^[2].
Methylene blue (Basic Blue 9) (2 mg/kg; i.v.; once, for 1 d; TBI-treated male BALB/c mice) reduces TBI-induced edema and neuroinflammation and reduces acute depression-like behavior^[3].
Methylene blue (Basic Blue 9) (2 mg/kg; i.v.; once, for 1 d; TBI-treated male BALB/c mice) reduces the percentage of inflammatory factor^[3].

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

Animal Model:	Male NMRI mice^[1]
Dosage:	50 and 100 mg/kg
Administration:	Intraperitoneal injection; once, for 25 minutes
Result:	Reduced the prepulse inhibition and reduced the increase in locomotor activity caused by phencyclidine (PCP).

Animal Model:	CaMKIIα-tTA transactivator mice^[2]
Dosage:	20 and 40 mg/kg
Administration:	Oral administration; daily, for 6 months
Result:	Inhibited Tau aggregation in CaMKIIα-tTA transactivator mice.

Animal Model:	TBI-treated male BALB/c mice^[3]
Dosage:	2 mg/kg
Administration:	Intravenous injection; once, for 1 day
Result:	Decreased the level of CD14, IL-1β, TNF-α, and CCL2 mRNA.

Animal Model:	TBI-treated male BALB/c mice^[3]
Dosage:	2 mg/kg
Administration:	Intravenous injection; once, for 1 day
Result:	Reduced the percentage of myeloid (CD11b+/GR1+) cells, reduced IL-1β and enhanced IL-10 expression in microglia.

Molecular Weight

319.85

Formula

C₁₆H₁₈ClN₃S

CAS No.

61-73-4

Appearance

Solid

Color

Brown to reddish brown

SMILES

CN(C)C1=CC=C2C(SC(C(C=C/3)=N2)=CC3=[N+](C)/C)=C1.[Cl-]

Shipping

Room temperature in continental US; may vary elsewhere.

Storage

RT, sealed storage, away from moisture and light

In solvent	-80°C	1 year
	-20°C	6 months

Solvent & Solubility

In Vitro:

H₂O : 50 mg/mL (156.32 mM; Need ultrasonic)

Preparing
Stock Solutions

Concentration Solvent Mass	1 mg	5 mg	10 mg

1 mM	3.1265 mL	15.6323 mL	31.2647 mL
5 mM	0.6253 mL	3.1265 mL	6.2529 mL
10 mM	0.3126 mL	1.5632 mL	3.1265 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, 1 year; -20°C, 6 months. When stored at -80°C, please use it within 1 year. When stored at -20°C, please use it within 6 months.

* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

Molarity Calculator
Dilution Calculator

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

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

This equation is commonly abbreviated as: C₁V₁ = C₂V₂

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In Vivo Dissolution Calculator

Please enter the basic information of animal experiments:

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Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.

Calculation results:

Working solution concentration: mg/mL

This product has good water solubility, please refer to the measured solubility data in water/PBS/Saline for details.

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).

Purity & Documentation

Purity: 95.05%

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Data Sheet (279 KB) SDS (393 KB)

COA (246 KB) HNMR (256 KB) RP-HPLC (224 KB) MS (68 KB)

Handling Instructions (2659 KB)

Dyeing Example

References

[1]. Klamer D, et, al. Phencyclidine-induced behaviour in mice prevented by methylene blue. Basic Clin Pharmacol Toxicol. 2004 Feb;94(2):65-72. [Content Brief]

[2]. Hochgräfe K, et, al. Preventive methylene blue treatment preserves cognition in mice expressing full-length pro-aggregant human Tau. Acta Neuropathol Commun. 2015 May 10;3:25. [Content Brief]

[3]. Fenn AM, et, al. Methylene blue attenuates traumatic brain injury-associated neuroinflammation and acute depressive-like behavior in mice. J Neurotrauma. 2015 Jan 15;32(2):127-38. [Content Brief]

[4]. Kalaitzakis D, et al. Methylene Blue as a Photosensitizer and Redox Agent: Synthesis of 5-Hydroxy-1H-pyrrol-2(5H)-ones from Furans. Angew Chem Int Ed Engl. 2015 May 18;54(21):6283-7. [Content Brief]

Complete Stock Solution Preparation Table

Optional Solvent	Concentration Solvent Mass	1 mg	5 mg	10 mg	25 mg

H₂O	1 mM	3.1265 mL	15.6323 mL	31.2647 mL	78.1616 mL
	5 mM	0.6253 mL	3.1265 mL	6.2529 mL	15.6323 mL
	10 mM	0.3126 mL	1.5632 mL	3.1265 mL	7.8162 mL
	15 mM	0.2084 mL	1.0422 mL	2.0843 mL	5.2108 mL
	20 mM	0.1563 mL	0.7816 mL	1.5632 mL	3.9081 mL
	25 mM	0.1251 mL	0.6253 mL	1.2506 mL	3.1265 mL
	30 mM	0.1042 mL	0.5211 mL	1.0422 mL	2.6054 mL
	40 mM	0.0782 mL	0.3908 mL	0.7816 mL	1.9540 mL
	50 mM	0.0625 mL	0.3126 mL	0.6253 mL	1.5632 mL
	60 mM	0.0521 mL	0.2605 mL	0.5211 mL	1.3027 mL
	80 mM	0.0391 mL	0.1954 mL	0.3908 mL	0.9770 mL
	100 mM	0.0313 mL	0.1563 mL	0.3126 mL	0.7816 mL

* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

Methylene Blue 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.

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Methylene Blue (Synonyms: Basic Blue 9; CI-52015; Methylthioninium chloride)

Customer Review

Other Forms of Methylene Blue:

Methylene Blue Related Antibodies

28 Publications Citing Use of MCE Methylene Blue

Featured Recommendations

•Related Small Molecules:

•Related Proteins:

View All Monoamine Oxidase Isoform Specific Products:

View All NO Synthase Isoform Specific Products:

Biological Activity

Purity & Documentation

References

Customer Review

Methylene Blue Related Antibodies

Molarity Calculator

Dilution Calculator

Complete Stock Solution Preparation Table

Methylene Blue Related Classifications

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