Monochlorobimane (Chlorobimane) 是一种荧光染料 (λex=380 nm,λem=470 nm),用于测定细胞中谷胱甘肽 (GSH)。
生物活性 | Monochlorobimane (Chlorobimane) is afluorescent dye(λex=380 nm, λem=470 nm) to measure glutathione (GSH) in cellular assays[1]. |
体外研究 (In Vitro) | 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 EX/EM= 380/465 ±20 nm respectively (blue).
|
分子量 | |
性状 | |
Formula | |
CAS 号 | |
Emission (Em) | Em 450-495 nm Blue |
Excitation (Ex) | Ex<380 nm Ultraviolet |
运输条件 | Room temperature in continental US; may vary elsewhere. |
储存方式 | -20°C, protect from light *In solvent : -80°C, 6 months; -20°C, 1 month (protect from light) |
溶解性数据 | In Vitro: DMSO : 50 mg/mL(220.59 mM;Need ultrasonic) 配制储备液 1 mM | 4.4119 mL | 22.0595 mL | 44.1189 mL | 5 mM | 0.8824 mL | 4.4119 mL | 8.8238 mL | 10 mM | 0.4412 mL | 2.2059 mL | 4.4119 mL |
*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。 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: 2.5 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。 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。 *以上所有助溶剂都可在本网站选购。
|
染色示例 | |