LPS 激活的 BV2 小胶质细胞可以改变其免疫学特征，并通过亚甲蓝 (Basic Blue 9)（4.5 μM；BV2 小胶质细胞）降低其 CD14、IL-1β、TNF-α 和 CCL2 mRNA 水平 [3]。

每 25 分钟一次以 50 和 100 mg/kg 的剂量腹膜内给予雄性 NMRI 小鼠一次亚甲蓝（Basic Blue 9）可减少前脉冲抑制的不足[1]。 ?在表达全长促聚集人类 Tau 能力的小鼠中，Basic Blue 9（20 和 40 mg/kg；口服；每天一次，持续 6 个月；CaMKIIα-tTA 反式激活小鼠）保留了认知能力[2]。 ?在经 TBI 治疗的雄性 BALB/c 小鼠中，Basic Blue 9（2 mg/kg；静脉注射；1 天一次）可减少最初的抑郁样行为以及 TBI 引起的水肿和神经炎症 [3]。 ?通过每天一次以 2 mg/kg 的剂量静脉注射亚甲蓝（Basic Blue 9），TBI 治疗的雄性 BALB/c 小鼠中炎症因子的百分比降低[3]。

Animal/Disease Models: Male NMRI mice [1]
Doses: 50 and 100 mg/kg
Route of Administration: intraperitoneal (ip) injection; once for 25 minutes
Experimental Results: diminished prepulse inhibition and diminished phencyclidine (PCP)-induced locomotor activity Increase.

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Animal/Disease Models: CaMKIIα-tTA transactivated mice [2]
Doses: 20 and 40 mg/kg
Route of Administration: Oral; one time/day for 6 months
Experimental Results: Inhibition of Tau aggregation in CaMKIIα-tTA transactivated mice .

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Animal/Disease Models: TBI-treated male balb/c (Bagg ALBino) mouse [3]
Doses: 2 mg/kg
Route of Administration: intravenous (iv) (iv)injection; once for 1 day
Experimental Results: CD14, IL-1β, TNF-α and CCL2 mRNA levels were diminished.

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Animal/Disease Models: TBI-treated male balb/c (Bagg ALBino) mouse [3]
Doses: 2 mg/kg
Route of Administration: intravenous (iv) (iv)injection; once for 1 day
Experimental Results: The percentage of bone marrow (CD11b+/GR1+) cells in microglia diminished , IL-1β was diminished and IL-10 expression was enhanced.

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

C16H18CLN3S

319.85222

61-73-4

Methylene blue trihydrate;7220-79-3;Methylene blue hydrate;122965-43-9

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

CI52015.; Methylene blue

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

H2O : ~50 mg/mL (~156.32 mM)

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*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.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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请根据您的实验动物和给药方式选择适当的溶解配方/方案：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
5、为保证最佳实验结果，工作液请现配现用！
6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。