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  3. Nile Blue A sulfate

Nile Blue A sulfate  (Synonyms: Nile blue sulfate)

Cat. No.: HY-101900 Purity: ≥98.0%
COA Handling Instructions

Nile Blue A (Nile blue sulfate) is used to differentiate melanins and lipofuscins. It is also useful for staining fats and preparation of an amperometric glucose sensor.

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

Nile Blue A sulfate Chemical Structure

Nile Blue A sulfate Chemical Structure

CAS No. : 3625-57-8

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10 mM * 1 mL in DMSO
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Customer Review

Based on 1 publication(s) in Google Scholar

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  • Biological Activity

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Description

Nile Blue A (Nile blue sulfate) is used to differentiate melanins and lipofuscins. It is also useful for staining fats and preparation of an amperometric glucose sensor[1].

In Vitro

Nile blue A is a basic oxazine dye which is soluble in water and ethyl alcohol. Nile blue A is a satisfactory stain for PHB granules in bacteria and is in fact superior to Sudan black B for this purpose. Poly-p3-hydroxybutyrate granules exhibits a strong orange fluorescence when stained with Nile blue A. Nile blue A appears to stain many more PHB granules than Sudan black B does and is not as easily ished from the cell by decolorization procedures[1]. Nile blue A is used as a stain for polyhydroxyalkanoic acid-accumulating microorganisms or to detect polyhydroxyalkanoic acids in microorganisms. Escherichia coli cells that do not accumulate detectable polyhydroxyalkanoic acids can be stained with Nile blue A and that this staining is sufficient for identifying these cells in fluorescence-activated cell sorting (FACS) experiments. Nile blue A staining does not affect either surface display of peptides or specific labeling of these peptides by a second fluorescence. Staining E. coli for flow cytometry using Nile blue A is an easy-to-handle and low-cost alternative to other fluorescent dyes or the intracellular expression of, for example, green fluorescent protein[2]. Nile blue A is one of the most studied benzophenoxazine dyes, as a potent photosensitizer for photodynamic therapy. The dye when administered intravenously disperses throughout the body by circulating through blood and is taken up by most cells that emphasize its interaction with various biomolecule[3].

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

Molecular Weight

366.42

Formula

C20H20N3O.1/2O4S2

CAS No.
Appearance

Solid

Color

Light brown to black

SMILES

CCN(C1=CC2=[O+]C3=C(C4=CC=CC=C4C(N)=C3)N=C2C=C1)CC.O=S([O-])([O-])=O.[0.5]

Shipping

Room temperature in continental US; may vary elsewhere.

Storage

4°C, sealed storage, away from moisture and light

*In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)

Solvent & Solubility
In Vitro: 

DMSO : ≥ 150 mg/mL (409.37 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 2.7291 mL 13.6455 mL 27.2911 mL
5 mM 0.5458 mL 2.7291 mL 5.4582 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 (sealed storage, away from moisture and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

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  • Dilution Calculator

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

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This equation is commonly abbreviated as: C1V1 = C2V2

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In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:

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Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Calculation results:
Working solution concentration: mg/mL
Purity & Documentation
References
Cell Assay
[1][2][3]

1% aqueous solution of Nile blue A is prepared and filtered before use. Mild heating may be necessary to fully dissolve the stain. Heat-fixed smears of bacterial cells are stained with the Nile blue A solution at 55°C for 10 min in a coplin staining jar. After being stained, the slides are washed with tap water to remove excess stain and with 8% aqueous acetic acid for 1 min. The stained smear is washed and blotted dry with bibulous paper, remoistened with tap water, and covered with a no. 1 glass cover slip. The preparation is examined with a Nikon Labphot microscope with an episcopic fluorescence attachment[1]. The PHA− strain Escherichia coli UT5600(DE3) is stained with Nile blue A. In an Erlenmeyer flask, 20 mL of Luria–Bertani (LB) broth is inoculated with one colony of UT5600(DE3) and incubated for 14 h at 37 °C and 200 rpm. Subsequently, 20 mL of LB broth containing Nile blue A in a final concentration of 0.5 μg/mL is inoculated with 200 μL of the 14-h culture and cultured to an optical density at 578 nm (OD578) of 0.6. As a control, 20 mL of LB broth (without Nile blue A) is inoculated with 200 μL of the 14-h culture and is also cultured to an optical density at OD578 of 0.6. Every 20 min, the OD578 is determined for both cultures to verify whether there is any influence of the dye on the growth of the bacteria[2]. Nile blue A stock solution is prepared using ethanol as the solvent. The concentration of NB is maintained at 5 μM for all the studies. The solutions are left for 1 h to achieve equilibrium before spectral measurements. The absorption spectra are recorded using Shimadzu Spectrophotometer (UV-1800) and the emission spectra are recorded using Jobin–Yvon Spectrofluorimeter. A 450 nm nano-LED is used as the light source and the fluorescence lifetime is collected at λem=672 nm[3].

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 (sealed storage, away from moisture and 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 2.7291 mL 13.6455 mL 27.2911 mL 68.2277 mL
5 mM 0.5458 mL 2.7291 mL 5.4582 mL 13.6455 mL
10 mM 0.2729 mL 1.3646 mL 2.7291 mL 6.8228 mL
15 mM 0.1819 mL 0.9097 mL 1.8194 mL 4.5485 mL
20 mM 0.1365 mL 0.6823 mL 1.3646 mL 3.4114 mL
25 mM 0.1092 mL 0.5458 mL 1.0916 mL 2.7291 mL
30 mM 0.0910 mL 0.4549 mL 0.9097 mL 2.2743 mL
40 mM 0.0682 mL 0.3411 mL 0.6823 mL 1.7057 mL
50 mM 0.0546 mL 0.2729 mL 0.5458 mL 1.3646 mL
60 mM 0.0455 mL 0.2274 mL 0.4549 mL 1.1371 mL
80 mM 0.0341 mL 0.1706 mL 0.3411 mL 0.8528 mL
100 mM 0.0273 mL 0.1365 mL 0.2729 mL 0.6823 mL
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    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.

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Nile Blue A sulfate
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