| Bioactivity | Monochlorobimane (Chlorobimane) is a fluorescent dye (λex=380 nm, λem=470 nm) to measure glutathione (GSH) in cellular assays[1]. Storage: protect from light. |
| Invitro | 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 protocol3.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 detection6.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). |
| Name | Monochlorobimane |
| CAS | 76421-73-3 |
| Formula | C10H11ClN2O2 |
| Molar Mass | 226.66 |
| Appearance | Solid |
| Transport | Room temperature in continental US; may vary elsewhere. |
| Storage | -20°C, protect from light *In solvent : -80°C, 6 months; -20°C, 1 month (protect from light) |
| Reference | [1]. Kamencic H, et al. Monochlorobimane fluorometric method to measure tissue glutathione. Anal Biochem. 2000 Nov 1;286(1):35-7. [2]. Manuela D. Machado, et al. Assessment of cellular reduced glutathione content in Pseudokirchneriella subcapitata using monochlorobimane. Journal of Applied Phycology volume 24, pages1509–1516(2012). |
Monochlorobimane
CAS: 76421-73-3 F: C10H11ClN2O2 W: 226.66
Monochlorobimane (Chlorobimane) is a fluorescent dye (λex=380 nm, λem=470 nm)to measure glutathione (GSH) in cellular
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