TMRE is a mitochondria specific dye (λ_ex=550 nm, λ_em=575 nm).

TMRE is a mitochondria specific dye (λ_ex=550 nm, λ_em=575 nm)^[1]. Multidirectional dynamic movement of TMRE is observed in epithelial cells and bidirectional dynamic movement is seen in the superficial cortical fiber cells of live bovine lenses. In the epithelium, the movement of TMRE fluorescence is up to 5 μm/min whereas in the superficial cortex the observed movement is up to 18.5 μm/min. The movement of TMRE fluorescence is abolished with treatment of the uncoupler, carbonyl cyanide m-chlorophenylhydrazone (CCCP)^[2].

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

514.95

C₂₆H₂₇ClN₂O₇

115532-52-0

Room temperature in continental US; may vary elsewhere.

4deg;C, sealed storage, away from moisture and light

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

In Vitro:;

DMSO : 27.78 mg/mL (53.95 mM; Need ultrasonic)

*

请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。
储备液的保存方式和期限：-80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)。-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。

In Vivo:

请根据您的实验动物和给药方式选择适当的溶解方案。以下溶解方案都请先按照 In Vitro 方式配制澄清的储备液，再依次添加助溶剂：

——为保证实验结果的可靠性，澄清的储备液可以根据储存条件，适当保存；体内实验的工作液，建议您现用现配，当天使用； 以下溶剂前显示的百
分比是指该溶剂在您配制终溶液中的体积占比；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的方式助溶

• 1.

  请依序添加每种溶剂：;10% DMSO ;; 40% PEG300 ;; 5% Tween-80 ;; 45% saline

  Solubility: ≥ 2.08 mg/mL (4.04 mM); Clear solution

  此方案可获得 ≥ 2.08 mg/mL (4.04 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 20.8 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀；向上述体系中加入50 μL Tween-80，混合均匀；然后继续加入 450 μL生理盐水定容至 1 mL。

  将 0.9 g 氯化钠，完全溶解于 100 mL ddH₂O 中，得到澄清透明的生理盐水溶液

• 2.

  请依序添加每种溶剂：;10% DMSO ;; 90% (20% SBE-β-CD in saline)

  Solubility: ≥ 2.08 mg/mL (4.04 mM); Clear solution

  此方案可获得 ≥ 2.08 mg/mL (4.04 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 20.8 mg/mL 的澄清 DMSO 储备液加到 900 μL 20% 的 SBE-β-CD 生理盐水水溶液中，混合均匀。

  将 2 g 磺丁基醚 β-环糊精加入 5 mL 生理盐水中，再用生理盐水定容至 10 mL，完全溶解，澄清透明

• [1]. Crowley LC, et al. Measuring Mitochondrial Transmembrane Potential by TMRE Staining. Cold Spring Harb Protoc. 2016 Dec 1;2016(12):pdb.prot087361.

  [2]. Bantseev V, et al. Confocal laser scanning microscopy imaging of dynamic TMRE movement in the mitochondria of epithelial and superficial cortical fiber cells of bovine lenses. Mol Vis. 2005 Jul 14;11:518-23.

The entire experiment should be performed at room temperature because temperature will directly impact mitochondrial transmembrane potential and TMRE staining. Cells should never be placed, centrifuged, incubated, or washed at 4°C or have ice-cold buffers or media added. Treat the cells with a cytotoxic stimulus. Harvest cells and resuspend at 5×10⁵ cells/mL in culture medium containing 150 nM TMRE. Incubate for 5 min at room temperature in the dark. Add Carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP) (5 μM final concentration) to an aliquot of untreated cells and incubate for 5 min at room temperature in the dark. Turn on the appropriate laser on the flow cytometer. Set up a histogram plot to detect TMRE using log scale^[1].

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

• [1]. Crowley LC, et al. Measuring Mitochondrial Transmembrane Potential by TMRE Staining. Cold Spring Harb Protoc. 2016 Dec 1;2016(12):pdb.prot087361.

  [2]. Bantseev V, et al. Confocal laser scanning microscopy imaging of dynamic TMRE movement in the mitochondria of epithelial and superficial cortical fiber cells of bovine lenses. Mol Vis. 2005 Jul 14;11:518-23.