MBD
Based on 1 Customer Validation
MBD (7-(p-Methoxybenzylamino)-4-nitrobenz-2,1,3-oxadiazole) is a hydrophobic fluorescent probe that is used to detect conformational changes of the complement subcomponent C1r during its autoactivation process. MBD acts by selectively binding to exposed hydrophobic regions on the C1r macromolecule. The detection wavelength of MBD in the free unbound state is Ex/Em = 490/560 nm, whereas the maximum emission wavelength of MBD upon binding to C1r exhibits a blue shift of approximately 20 nm relative to 560 nm.
For research use only. We do not sell to patients.
- Purity : 99.60%
- CAS No.: 33984-50-8
- Formula: C14H12N4O4
- Molecular Weight:300.27
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Storage:
4°C, protect from light
* In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)
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Tel: 609-228-6898 E-mail: [email protected] [email protected]
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Biological Activity
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Chemical Information
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Solvent & Solubility
- Purity & Documentation
- References
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Help & FAQs
Biological Activity
Description
In Vitro
Guidelines (The following is our recommended experimental protocol. This protocol is for reference only and should be modified according to your specific needs).
1. Stock Solution Preparation[2]
1.1 Solvent: DMSO.
2. Working Solution Preparation
2.1 Diluent: 20 mM Tris-HCl buffer containing 0.15 M NaCl.
2.2 Working concentration: The MBD concentration in the final assay system is 4 μM.
2.3 Note: Adjust the working solution concentration as needed; prepare fresh before use.
3. Staining Procedure
3.1 Sample type: Purified C1r protein sample at a concentration of approximately 40 μg protein/mL.
3.2 Incubation conditions: Perform the binding assay using 4 μM MBD working solution at 20°C and pH 7.4.
5. Detection and Analysis
5.1 Instrument: Spectrofluorometer.
5.1.1 Excitation/emission wavelengths: Set the excitation wavelength to 490 nm and record the fluorescence emission spectrum in the range of 500-600 nm.
5.2 Result analysis:
5.2.1 Free MBD exhibits maximum fluorescence emission at 560 nm; upon addition of proenzyme C1r, the fluorescence intensity increases significantly, and the maximum emission wavelength shifts by approximately 20 nm toward shorter wavelengths.
5.2.2 Upon addition of activated C1r, the fluorescence spectrum of MBD shows no significant change, indicating that proteolytic activation disrupts the hydrophobic region in proenzyme C1r that binds MBD.
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. .
Chemical Information
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CAS No. 33984-50-8
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Appearance Solid
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Molecular Weight 300.27
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Formula C14H12N4O4
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Color Orange to red
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SMILES
O=[N+](C1=CC=C(NCC2=CC=C(OC)C=C2)C3=NON=C31)[O-]
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Synonyms
7-(p-Methoxybenzylamino)-4-nitrobenz-2,1,3-oxadiazole
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Shipping
Room temperature in continental US; may vary elsewhere.
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Storage
4°C, protect from light
* In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)
Solvent & Solubility
In Vitro:
DMSO : 25 mg/mL (83.26 mM; ultrasonic and warming and heat to 60°C; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)
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.
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.
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
Purity & Documentation
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Data Sheet (282 KB)
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SDS (252 KB)
- English - EN (252 KB)
- Français - FR (252 KB)
- Deutsch - DE (252 KB)
- Norwegian - NO (252 KB)
- Español - ES (252 KB)
- Swedish - SV (252 KB)
- Italian - IT (252 KB)
- Korean - KR (252 KB)
- Portuguese - PT (252 KB)
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Handling Instructions (2659 KB)
References
[1]. Kenner RA, et al. A new fluorescent probe for protein and nucleoprotein conformation. Binding of 7-(p-methoxybenzylamino)-4-nitrobenzoxadiazole to bovine trypsinogen and bacterial ribosomes. Biochemistry. 1971 Nov 23;10(24):4433-40. [Content Brief]
[2]. Kasahara Y, et al. Formation of a conformationally changed C1r, a subcomponent of the first component of human complement, as an intermediate of its autoactivation reaction. FEBS Lett. 1982 May 3;141(1):128-31. [Content Brief]
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 | 3.3303 mL | 16.6517 mL | 33.3034 mL | 83.2584 mL |
| 5 mM | 0.6661 mL | 3.3303 mL | 6.6607 mL | 16.6517 mL | |
| 10 mM | 0.3330 mL | 1.6652 mL | 3.3303 mL | 8.3258 mL | |
| 15 mM | 0.2220 mL | 1.1101 mL | 2.2202 mL | 5.5506 mL | |
| 20 mM | 0.1665 mL | 0.8326 mL | 1.6652 mL | 4.1629 mL | |
| 25 mM | 0.1332 mL | 0.6661 mL | 1.3321 mL | 3.3303 mL | |
| 30 mM | 0.1110 mL | 0.5551 mL | 1.1101 mL | 2.7753 mL | |
| 40 mM | 0.0833 mL | 0.4163 mL | 0.8326 mL | 2.0815 mL | |
| 50 mM | 0.0666 mL | 0.3330 mL | 0.6661 mL | 1.6652 mL | |
| 60 mM | 0.0555 mL | 0.2775 mL | 0.5551 mL | 1.3876 mL | |
| 80 mM | 0.0416 mL | 0.2081 mL | 0.4163 mL | 1.0407 mL |
MBD Related Classifications
HY-141667 Related Classifications
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Keywords
- MBD
- 33984-50-8
- 7-(p-Methoxybenzylamino)-4-nitrobenz-2,1,3-oxadiazole
- Fluorescent Dye
- complement subcomponent C1r
- conformational changes
- hydrophobic fluorescent probe
- autoactivation
- Tris-HCl buffer
- fluorescent emission spectra
- zymogen C1r
- proteolytically activated two-chain form of C1r
- hydrophobic binding pocket
- proteolytic activation
- Inhibitor
- inhibitor
- inhibit