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  4. Monochlorobimane

Monochlorobimane  (Synonyms: Chlorobimane，mBBr)

Cat. No.: HY-101899 Purity: 99.86%
COA Handling Instructions

Molarity Calculator

Monochlorobimane (Chlorobimane) is a fluorescent dye (λex=380 nm, λem=470 nm) to measure glutathione (GSH) in cellular assays.

For research use only. We do not sell to patients.

Monochlorobimane Chemical Structure

Monochlorobimane Chemical Structure

CAS No. : 76421-73-3

Size Price Stock Quantity
1 mg USD 30 In-stock
5 mg USD 60 In-stock
10 mg USD 107 In-stock
25 mg USD 224 In-stock
50 mg USD 380 In-stock
100 mg USD 650 In-stock
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Customer Review

Based on 4 publication(s) in Google Scholar

Monochlorobimane Related Antibodies

Top Publications Citing Use of Products

  • Biological Activity

  • Protocol

  • Purity & Documentation

  • References

  • Customer Review

Description

Monochlorobimane (Chlorobimane) is a fluorescent dye (λex=380 nm, λem=470 nm) to measure glutathione (GSH) in cellular assays[1].

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).

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Monochlorobimane Related Antibodies
Molecular Weight

226.66

Formula

C10H11ClN2O2
CAS No.
Appearance

Solid

Color

Light yellow to yellow

Emission (Em)

490
Excitation (Ex)

395
SMILES

O=C1N(C(C(C)=C2C)=O)N2C(CCl)=C1C
Shipping

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)

Solvent & Solubility
In Vitro: 

DMSO : 50 mg/mL (220.59 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

Preparing
Stock Solutions
Concentration Solvent Mass 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
View the Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

  • Molarity Calculator

  • Dilution Calculator

Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

Mass
=
Concentration
×
Volume
×
Molecular Weight *

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

This equation is commonly abbreviated as: C1V1 = C2V2

Concentration (start)

C1

×
Volume (start)

V1

=
Concentration (final)

C2

×
Volume (final)

V2

In Vivo:

Select the appropriate dissolution method based on your experimental animal and administration route.

For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

  • Protocol 1

    Add each solvent one by one:  10% DMSO    40% PEG300    5% Tween-80    45% Saline

    Solubility: 2.5 mg/mL (11.03 mM); Clear solution; Need ultrasonic

    This protocol yields a clear solution of 2.5 mg/mL.

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.

    Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
  • Protocol 2

    Add each solvent one by one:  10% DMSO    90% (20% SBE-β-CD in Saline)

    Solubility: 2.5 mg/mL (11.03 mM); Suspended solution; Need ultrasonic

    This protocol yields a suspended solution of 2.5 mg/mL. Suspended solution can be used for oral and intraperitoneal injection.

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.

    Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.
In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:

Dosage

mg/kg

Animal weight
(per animal)

g

Dosing volume
(per animal)

μL

Number of animals

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
%
DMSO +
% 
+
%
Tween-80 +
%
Saline
Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).
Calculation results:
Working solution concentration: mg/mL
Method for preparing stock solution: mg drug dissolved in μL  DMSO (Stock solution concentration: mg/mL).

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only. If necessary, please contact MedChemExpress (MCE).
Method for preparing in vivo working solution for animal experiments: Take μL DMSO stock solution, add μL . μL , mix evenly, next add μL Tween 80, mix evenly, then add μL Saline.
 If the continuous dosing period exceeds half a month, please choose this protocol carefully.
Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
Purity & Documentation

Purity: 99.86%

COA (256 KB) HNMR (216 KB) LCMS (95 KB)

Handling Instructions (2659 KB)
Dyeing Example
References
Kinase Assay
[1]

Monochlorobimane (mCB) is added to the second half liver tissue homogenate to a final concentration of 100 mM along with glutathione S-transferase (1 U/mL) obtained from equine liver; the homogenate is then allowed to incubate at room temperature for 30 min. The Glutathione (GSH)-Monochlorobimane adduct is measured in a microtiter reader[1].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.
References

Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
DMSO 1 mM 4.4119 mL 22.0595 mL 44.1189 mL 110.2974 mL
5 mM 0.8824 mL 4.4119 mL 8.8238 mL 22.0595 mL
10 mM 0.4412 mL 2.2059 mL 4.4119 mL 11.0297 mL
15 mM 0.2941 mL 1.4706 mL 2.9413 mL 7.3532 mL
20 mM 0.2206 mL 1.1030 mL 2.2059 mL 5.5149 mL
25 mM 0.1765 mL 0.8824 mL 1.7648 mL 4.4119 mL
30 mM 0.1471 mL 0.7353 mL 1.4706 mL 3.6766 mL
40 mM 0.1103 mL 0.5515 mL 1.1030 mL 2.7574 mL
50 mM 0.0882 mL 0.4412 mL 0.8824 mL 2.2059 mL
60 mM 0.0735 mL 0.3677 mL 0.7353 mL 1.8383 mL
80 mM 0.0551 mL 0.2757 mL 0.5515 mL 1.3787 mL
100 mM 0.0441 mL 0.2206 mL 0.4412 mL 1.1030 mL
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Monochlorobimane Related Classifications

Help & FAQs
  • Do most proteins show cross-species activity?

    Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

Keywords:

Monochlorobimane76421-73-3Chlorobimane，mBBrFluorescent DyeInhibitorinhibitorinhibit

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