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Nitro blue tetrazolium chloride  (Synonyms: NBT)

Cat. No.: HY-15925 Purity: 99.82%
COA
SDS
Handling Instructions Technical Support

Nitro blue tetrazolium chloride (NBT) is a substrate for dehydrogenases; is used with the alkaline phosphatase substrate 5-Bromo-4-Chloro-3-Indolyl Phosphate (BCIP) in western blotting and immunohistological staining procedures.

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

CAS No. : 298-83-9

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Based on 6 publication(s) in Google Scholar

Nitro blue tetrazolium chloride Related Antibodies

Top Publications Citing Use of Products

    Nitro blue tetrazolium chloride purchased from MedChemExpress. Usage Cited in: Eur J Med Chem. 2024 Aug 13:277:116779.  [Abstract]

    The Nitro blue tetrazolium chloride (NBT, 0.25 mg/mL; 30 min) test of NB4 cells treated with DMSO, ATRA (0.5 μM), 5a (0.5 μM) or 5b (0.5 μM) for 6 days. The percentage of NBT positive cells was shown. Scale bar, 56.3 μm.

    Nitro blue tetrazolium chloride purchased from MedChemExpress. Usage Cited in: Preprints. 2024 Sep 9.

    Ten-day old seedlings were chilling and freezing treated 3 or 6 h, Leaves were then incubated in an Nitro blue tetrazolium chloride (NBT) solution (0.1% NBT in 10 mM sodium azide and 10 mM phosphate buffer, pH 7.8) overnight. The phenotype was photographed, and the activity of SOD, CAT, POD, O2-, MDA, soluble sugars, and proline content was detected. The phenotype of ROS accumulation.

    Nitro blue tetrazolium chloride purchased from MedChemExpress. Usage Cited in: J Plant Physiol. 2023 Sep:288:154076.

    ROS accumulation in BrEBF1 transgenic plants was detected with Nitro blue tetrazolium chloride (NBT) staining. The leaves were infiltrated with nitroblue tetrazolium (NBT) (0.1% NBT in 10 mM sodium azide and 10 mM phosphate buffer, pH 7.8) overnight.

    Nitro blue tetrazolium chloride purchased from MedChemExpress. Usage Cited in: Cell Death Dis. 2021 May 8;12(5):462.  [Abstract]

    H322 cells (1 × 104) with or without FTO depletion by two different shRNAs (shFTO-1 and shFTO-2) were cultured in soft agar for 3 weeks and then were stained with Nitro blue tetrazolium chloride solution (200 μL/ well) and incubated overnight at 37 °C. Cell clones were stained and counted from six different fields under a microscope. Scale bar: 100 μm.

    • Biological Activity

    • Purity & Documentation

    • References

    • Customer Review

    Description

    Nitro blue tetrazolium chloride (NBT) is a substrate for dehydrogenases; is used with the alkaline phosphatase substrate 5-Bromo-4-Chloro-3-Indolyl Phosphate (BCIP) in western blotting and immunohistological staining procedures[1].

    In Vitro

    Guidelines (The following is our recommended experimental protocol. This protocol is for reference only, and specific operations should be adjusted according to your actual needs).
    NBT and BCIP staining protocol
    1. Materials
    Dimethylformamide (DMF, HY-Y0345).
    Nitro blue tetrazolium chloride (NBT, HY-15925) stock solution: Dissolve 0.5 g NBT in 10 mL 70% aqueous DMF.
    BCIP stock solution: Dissolve 0.5 g BCIP in 10 mL 100% DMF.
    Substrate Buffer: 100 mM diethanolamine buffer (pH 9.5) containing 100 mM NaCl and 5 mM MgCl2.
    2. Methods
    2.1 To prepare the NBT/BCIP solution, add 66 μL NBT stock solution to 10 mL Substrate Buffer and mix thoroughly. Add 33 μL BCIP stock solution to the mixture. Use the NBT/BCIP solution within 1 hour.
    2.2 Place the blot into an appropriate container and add the NBT/BCIP solution. Use 10 mL solution for a 15 × 15 cm2 membrane.
    Agitate at room temperature and incubate the blot until the bands develop to the desired darkness. Typically, about 30 minutes is required.
    2.3 Remove the NBT/BCIP solution and rinse the blot with water to stop the reaction.

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

    Nitro blue tetrazolium chloride Related Antibodies
    Molecular Weight

    817.64

    Formula

    C40H30Cl2N10O6
    CAS No.
    Appearance

    Solid

    Color

    Light yellow to yellow

    SMILES

    COC1=CC(C2=CC=C(N3N=C(C4=CC=CC=C4)N=[N+]3C5=CC=C([N+]([O-])=O)C=C5)C(OC)=C2)=CC=C1N6N=C(C7=CC=CC=C7)N=[N+]6C8=CC=C([N+]([O-])=O)C=C8.[Cl-].[Cl-]
    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: 

    70%DMF : 100 mg/mL (122.30 mM; Need ultrasonic)

    H2O : 8 mg/mL (9.78 mM; Need ultrasonic)

    DMSO : 5 mg/mL (6.12 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)

    Preparing
    Stock Solutions
    Concentration Solvent Mass 1 mg 5 mg 10 mg
    1 mM 1.2230 mL 6.1152 mL 12.2303 mL
    5 mM 0.2446 mL 1.2230 mL 2.4461 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.

    * Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

    • Molarity Calculator

    • Dilution Calculator

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

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

    This equation is commonly abbreviated as: C1V1 = C2V2

    Concentration (start)

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

    Dosage

    mg/kg

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    (per animal)

    g

    Dosing volume
    (per animal)

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    Number of animals

    Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
    Calculation results:
    Working solution concentration: mg/mL
    Purity & Documentation

    Purity: 99.82%

    SDS (560 KB)

    COA (259 KB) HNMR (195 KB) LCMS (120 KB)

    Handling Instructions (2659 KB)
    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 / H2O / 70%DMF 1 mM 1.2230 mL 6.1152 mL 12.2303 mL 30.5758 mL
    5 mM 0.2446 mL 1.2230 mL 2.4461 mL 6.1152 mL
    70%DMF 10 mM 0.1223 mL 0.6115 mL 1.2230 mL 3.0576 mL
    15 mM 0.0815 mL 0.4077 mL 0.8154 mL 2.0384 mL
    20 mM 0.0612 mL 0.3058 mL 0.6115 mL 1.5288 mL
    25 mM 0.0489 mL 0.2446 mL 0.4892 mL 1.2230 mL
    30 mM 0.0408 mL 0.2038 mL 0.4077 mL 1.0192 mL
    40 mM 0.0306 mL 0.1529 mL 0.3058 mL 0.7644 mL
    50 mM 0.0245 mL 0.1223 mL 0.2446 mL 0.6115 mL
    60 mM 0.0204 mL 0.1019 mL 0.2038 mL 0.5096 mL
    80 mM 0.0153 mL 0.0764 mL 0.1529 mL 0.3822 mL
    100 mM 0.0122 mL 0.0612 mL 0.1223 mL 0.3058 mL

    * Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

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    Nitro blue tetrazolium chloride 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:

    Nitro blue tetrazolium298-83-9NBTFluorescent DyedehydrogenasesBCIPwestern blottingimmunohistologicalInhibitorinhibitorinhibit

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