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Monochlorobimane

CAS No. : 76421-73-3

MCE 站：Monochlorobimane

产品活性：Monochlorobimane (Chlorobimane) 是一种荧光染料 (λ_ex=380 nm，λ_em=470 nm)，用于测定细胞中谷胱甘肽 (GSH)。

研究领域：Others

作用靶点：Others

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 ?M 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 ?L 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 ?L of the culture suspension into the white opaque plate and record fluorescence at E_X/E_M= 380/465 ±20 nm respectively (blue).

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