Monochlorobimane (Chlorobimane) is afluorescent dye(λ_ex=380 nm, λ_em=470 nm) to measure glutathione (GSH) in cellular assays^[1].

Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
1. Preparation of control and experimental wells:
    The experiment should consist of parallel negative, positive and experimental wells respectively.
    Experimental wells: add 200 μL of cell culture media containing your GSH effector of interest at desired concentration (e.g. 200 μM H₂O₂)
2. Incubate the plate overnight at 37 ℃, 5 % CO₂. The incubation time varies depended on your normal protocol
3. Monochlorobimane (mBCl) is added to the cells at a final concentration of20-100 μMfrom a working solution of 1 mM. The stock solution of mBCl (50 mM) was prepared in dimethyl sulphoxide (DMSO) and stored at -20℃; the working solution was prepared before use by diluting the stock solution in 0.1 M PBS buffer (pH 7.0). In the assay the final concentration of DMSO was below 0.2% (v/v)
4. The cell suspensions were incubated with mBCl in the dark at 25℃ for ~2 h
5. Remove the dye and treatment by centrifugation at 700 X g for 5 min. Add 200 μL of the Assay Buffer to each well and continue to the preferred method of detection
6. Detection of intracellular GSH:
    Fluorescence plate reader: For adherent cells; read fluorescence directly off the culturing plate. If working with suspension cells; aliquot 200 μL of the culture suspension into the white opaque plate and record fluorescence at E_X/E_M= 380/465 ±20 nm respectively (blue).

226.66

Solid

C₁₀H₁₁ClN₂O₂

76421-73-3

Room temperature in continental US; may vary elsewhere.

-20°C, protect from light

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

DMSO : 50 mg/mL(220.59 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℃, 6 months; -20℃, 1 month (protect from light)。-80℃ 储存时，请在 6 个月内使用，-20℃ 储存时，请在 1 个月内使用。

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: 2.5 mg/mL (11.03 mM); Clear solution; Need ultrasonic

  此方案可获得 2.5 mg/mL (11.03 mM) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 25.0 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-CDin saline)

  Solubility: 2.5 mg/mL (11.03 mM); Suspended solution; Need ultrasonic

  此方案可获得 2.5 mg/mL (11.03 mM) 的均匀悬浊液，悬浊液可用于口服和腹腔注射。

  以 1 mL 工作液为例，取 100 μL 25.0 mg/mL 的澄清 DMSO 储备液加到 900 μL20% 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

• 
  Description: Monochlorobimane (Chlorobimane) can be used to detect intracellular GSH levels.
  Method: For tissue labeling.
  1. Stain frozen sections of eyeballs with Monochlorobimane (30 min).
  2. Rinse and observe staining tissue under a fluorescence microscope.