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| 5mg |
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| 10mg |
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| 25mg |
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| Targets |
Monochlorobimane targets glutathione (GSH), a key intracellular antioxidant molecule. It reacts with GSH thiols to form a fluorescent adduct. It is used for detecting and quantifying intracellular glutathione levels. Once bound, it emits high fluorescence, allowing for the tracking of GSH distribution and activity in live cells.
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| ln Vitro |
Advice (The procedure we suggest is as follows. This protocol should be adjusted to suit your unique requirements; it merely offers guidelines. 1. Setting up the experimental and control wells: Parallel negative, positive, and experimental wells should make up the experiment, in that order. Test wells: Pour in 200 μL of cell culture media with the appropriate amount of GSH effectors (e.g., 200 μM H2O2). 2. Let the plate sit in 5% CO2 at 37°C for the entire night. Depending on your standard procedure, incubation times can differ. 3. At a final concentration of 20–100 μM, add monochlorodimethane (mBCl) from the 1 mM working solution to the cells. Dimethyl sulfoxide (DMSO) was used to prepare stock solutions of mBCl (50 mM), which were then kept at -20°C. Prior to use, the stock solution was diluted in 0.1 M PBS buffer (pH 7.0). In the experiment, the final DMSO concentration is less than 0.2% (v/v). 4. For around two hours, incubate the cell suspension with mBCl at 25°C in the dark. 5. Centrifuge the dye at 700 X g for five minutes to remove it. Fill each well with 200 μL of Assay Buffer, then proceed with your chosen detection technique. 6. GSH detection inside cells: The fluorescence plate reader can read fluorescence straight from the culture plate and is appropriate for adherent cells. To record fluorescence (blue) at EX/EM = 380/465 ±20 nm, respectively, aliquot 200 μL of culture suspension into white opaque plates if employing suspension cells.
In vitro, Monochlorobimane is a non-fluorescent, cell-permeable thiol-reactive probe. It reacts with glutathione (GSH) to form a fluorescent adduct. It is widely used to measure intracellular reduced glutathione (GSH) levels. It has excitation (λex) at 380 nm and emission (λem) at 470 nm. |
| ln Vivo |
Monochlorobimane is not administered as a therapeutic agent in vivo. Its utility is limited to in vitro applications as a fluorescent probe for GSH detection.
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| Enzyme Assay |
In vitro assays with Monochlorobimane involve incubating cells with the probe (typically 1-100 µM) for 15-60 minutes. Cells are then washed and fluorescence is measured using a fluorescence microscope, flow cytometry, or a fluorescence plate reader with excitation at 380 nm and emission at 470 nm. The fluorescence intensity is proportional to the intracellular GSH level.
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| Cell Assay |
Cellular assays for Monochlorobimane are performed using various cell types. Cells are incubated with the probe at concentrations ranging from 1 to 100 µM for 15-60 minutes. After washing, fluorescence is measured by flow cytometry or fluorescence microscopy. The probe can be used to study GSH dynamics in response to oxidative stress, drug treatment, or genetic manipulations.
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| Animal Protocol |
Monochlorobimane is not used in in vivo animal studies as a therapeutic agent. It may be used in ex vivo applications to measure GSH levels in tissues harvested from animal studies.
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| ADME/Pharmacokinetics |
Metabolism / Metabolites
The known metabolites of monochlorodimaleic acid include bismaleic acid-glutathione. Monochlorobimane is a fluorescent probe and does not have pharmacokinetic properties as a drug. |
| Toxicity/Toxicokinetics |
Monochlorobimane is a fluorescent probe and is not intended for human consumption. Standard laboratory safety precautions should be followed.
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| References | |
| Additional Infomation |
Monochlorodimaleic acid is an organochlorine compound belonging to the pyrazolopyrazole class. It can be used as a fluorescent dye.
Monochlorobimane (mBCl) is a cell-permeable, non-fluorescent probe that reacts with glutathione (GSH) to form a fluorescent adduct. It is used to measure intracellular GSH levels. It has excitation at 380 nm and emission at 470 nm. It has a molecular weight of 226.66 and formula C10H11ClN2O2. It is not a drug. |
| Molecular Formula |
C10H11N2O2CL
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|---|---|
| Molecular Weight |
226.65954
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| Exact Mass |
226.051
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| CAS # |
76421-73-3
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| PubChem CID |
114886
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| Appearance |
Light yellow to yellow solid powder
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| Density |
1.4g/cm3
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| Boiling Point |
314.9ºC at 760 mmHg
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| Melting Point |
135-136ºC(lit.)
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| Flash Point |
144.2ºC
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| Index of Refraction |
1.61
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| LogP |
0.861
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
3
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| Rotatable Bond Count |
1
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| Heavy Atom Count |
15
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| Complexity |
437
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
SUIPVTCEECPFIB-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C10H11ClN2O2/c1-5-7(3)12-8(4-11)6(2)10(15)13(12)9(5)14/h4H2,1-3H3
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| Chemical Name |
7-(chloromethyl)-1,2,6-trimethylpyrazolo[1,2-a]pyrazole-3,5-dione
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: This product requires protection from light (avoid light exposure) during transportation and storage. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : ~50 mg/mL (~220.59 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: 2.5 mg/mL (11.03 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: 2.5 mg/mL (11.03 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 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 |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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