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> NBD-Cl [10199-89-0]

NBD-Cl [10199-89-0]

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Référence HY-D0042-1g

Conditionnement : 1g

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NBD-Cl is a nonfluorescent reagent which becomes highly fluorescent after reaction with thiol or amino groups.

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NBD-Cl Chemical Structure

NBD-Cl Chemical Structure

CAS No. : 10199-89-0

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Based on 1 publication(s) in Google Scholar

Description

NBD-Cl is a nonfluorescent reagent which becomes highly fluorescent after reaction with thiol or amino groups[1].

In Vitro

NBD-Cl (NBD chloride) forms highly fluorescent derivatives for detection of all the protein amino acids. In addition, NBD-Cl provides a simple and sensitive method for determination of N-terminal amino acids. The differences in intensity and color of fluorescence can be used to advantage to identify prolyl peptides[1].

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

NBD-Cl Related Antibodies
Masse moléculaire

199.55

Formule

C6H2ClN3O3
CAS No.

10199-89-0
Appearance

Solid

Color

Light yellow to yellow

Emission (Em)

515
Excitation (Ex)

498
SMILES

O=[N+](C1=CC=C(Cl)C2=NON=C21)[O-]
Livraison

Room temperature in continental US; may vary elsewhere.
Stockage

4°C, protect from light

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

Solvant et solubilité
In Vitro: 

DMSO : ≥ 100 mg/mL (501.13 mM; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

H2O : < 0.1 mg/mL (insoluble)

*"≥" means soluble, but saturation unknown.

Preparing
Stock Solutions
Concentration Solvent Mass 1 mg 5 mg 10 mg
1 mM 5.0113 mL 25.0564 mL 50.1128 mL
5 mM 1.0023 mL 5.0113 mL 10.0226 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.

  • Calculateur de molarité

  • Calculateur de dilution

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    90% (20% SBE-β-CD in Saline)

    Solubility: ≥ 2.5 mg/mL (12.53 mM); Clear solution

    This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

    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.
Pureté et documentation

Purity: 99.14%

COA (253 KB) HNMR (246 KB) RP-HPLC (86 KB)

Instruction de manipulation (2659 KB)
Références

  • [1]. Fager RS, et al. The use of NBD chloride (7 chloro-4-nitrobenzo-2-oxa-1,3-diazole) in detecting amino acids and as an N-terminal reagent. Anal Biochem. 1973 May;53(1):290-4.  [Content Brief]

Essai de kinase
[1]

Fluorescence Measurements[1]
Fluorescence spectra are recorded on a Hitachi MPF-24 Fluorescence Spectrophotometer. The emission spectrum is determined at the optimal wavelength of 464 nm. Thin-layer chromatography is carried out on Brinkmann glass supported analytical silica G thin-layer plates without fluorescent indicator. The NBD derivatives are visualized on thin-layer plates either by examination under an ultraviolet lamp with a long wavelength filter or by using the same lamp with a 2×2-in. Corning 5-57 filter and viewing the spots through a 2×2-in. Corning 3-68 filter. The former method produces a more intense fluorescence, but the latter produces a much greater contrast appearing as white spots against a dark background. NBD-Cl (NBD chloride) is allowed to react with amino acids and peptides under the conditions. Into 3 mL Pyrex test tubes are pipette the following: (1) 2.0 mL of ethanol, (2) 0.1 mL NBD-Cl solution in ethanol 1.41 pg/mL, (3) 0.1 mL ethanol saturated with sodium acetate, (4) 0.1 mL of amino acid solution 400-600μg/mL, and the mixture incubated at 75°C for 20 min. For reaction with peptides, 1.5 mL of ethanol, 0.1 mL ethanolic sodium acetate, 0.4 mL ethanolic NBD-Cl, 1.41 μg/mL, are added to 0.4 ml of the peptide and incubated under the same conditions.

MCE n'a pas confirmé de manière indépendante l'exactitude de ces méthodes. Ils sont pour référence seulement.
Références

  • [1]. Fager RS, et al. The use of NBD chloride (7 chloro-4-nitrobenzo-2-oxa-1,3-diazole) in detecting amino acids and as an N-terminal reagent. Anal Biochem. 1973 May;53(1):290-4.  [Content Brief]

  • [1]. Fager RS, et al. The use of NBD chloride (7 chloro-4-nitrobenzo-2-oxa-1,3-diazole) in detecting amino acids and as an N-terminal reagent. Anal Biochem. 1973 May;53(1):290-4.

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 5.0113 mL 25.0564 mL 50.1128 mL 125.2819 mL
5 mM 1.0023 mL 5.0113 mL 10.0226 mL 25.0564 mL
10 mM 0.5011 mL 2.5056 mL 5.0113 mL 12.5282 mL
15 mM 0.3341 mL 1.6704 mL 3.3409 mL 8.3521 mL
20 mM 0.2506 mL 1.2528 mL 2.5056 mL 6.2641 mL
25 mM 0.2005 mL 1.0023 mL 2.0045 mL 5.0113 mL
30 mM 0.1670 mL 0.8352 mL 1.6704 mL 4.1761 mL
40 mM 0.1253 mL 0.6264 mL 1.2528 mL 3.1320 mL
50 mM 0.1002 mL 0.5011 mL 1.0023 mL 2.5056 mL
60 mM 0.0835 mL 0.4176 mL 0.8352 mL 2.0880 mL
80 mM 0.0626 mL 0.3132 mL 0.6264 mL 1.5660 mL
100 mM 0.0501 mL 0.2506 mL 0.5011 mL 1.2528 mL
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NBD-Cl Related Classifications

  • Others
  • Others
  • Fluorescent Dye
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:

NBD-Cl10199-89-0NBDFluorescent DyeInhibitorinhibitorinhibit

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