耗材描述
Monochlorobimane
产品活性: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)。
产品来源: https://www.medchemexpress.cn/Monochlorobimane.html
研究领域:Others
作用靶点:Fluorescent Dye
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?°C, 5?% CO2. The incubation time varies depended on your normal protocol
3. Monochlorobimane (mBCl) is added to the cells at a final concentration of 20-100 μM from a working solution of 1 mM. The stock solution of mBCl (50 mM) was prepared in dimethyl sulphoxide (DMSO) and stored at -20°C; 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°C 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).
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