Nitro blue tetrazolium chloride (Synonyms: NBT)

Name: Nitro blue tetrazolium chloride
Brand: MedChemExpress
SKU: HY-15925
Rating: 4.5 (4 reviews)

Cat. No.: HY-15925 Purity: 99.82%: FAQs
Data Sheet SDS COA Handling Instructions

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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.

Nitro blue tetrazolium chloride Chemical Structure

CAS No. : 298-83-9

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Solid + Solvent
10 mM * 1 mL in DMSO ready for reconstitution	USD 33	In-stock	Estimated Time of Arrival: December 31
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10 mM * 1 mL in DMSO	USD 33	In-stock	Estimated Time of Arrival: December 31
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250 mg	USD 30	In-stock	Estimated Time of Arrival: December 31
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Based on 4 publication(s) in Google Scholar

4 Publications Citing Use of MCE Nitro blue tetrazolium chloride

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Description

In Vitro

Protocol for Staining with NBT and BCIP
1.Materials
Dimethylformamide (DMF, HY-Y0345).
Nitro blue tetrazolium chloride (NBT, HY-15925) Stock Solution: Dissolve 0.5 g of NBT in 10 mL of 70% DMF in water.
BCIP Stock Solution: Dissolve 0.5 g of BCIP in 10 mL 100% DMF.
Substrate Buffer: 100 mM diethanolamine buffer (pH 9.5) containing 100 mM NaCl and 5 mM MgCl₂.
2.Method
2.1 To prepare the NBT/BCIP solution, add 66 µL of stock NBT to 10 mL of substrate Buffer and mix well. Add 33 µL of stock BCIP to the solution. Use the NBT/BCIP solution within 1 hour.
2.2Place blot in a suitable container and add the NBT/BCIP Solution. Use 10 mL of solution for a 15 × 15cm² membrane.
Incubate blot at room temperature with agitation until the bands are suitably dark. Typically, ~30 minutes is required for development.
2.3 Remove the NBT/BCIP Solution and rinse blot with water to stop the reaction.

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

Molecular Weight

817.64

Formula

C₄₀H₃₀Cl₂N₁₀O₆

CAS No.

298-83-9

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
10 mM	0.1223 mL	0.6115 mL	1.2230 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.

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Dilution Calculator

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

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

This equation is commonly abbreviated as: C₁V₁ = C₂V₂

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 (3.06 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 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 (3.06 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.
Protocol 3

Add each solvent one by one: 10% DMSO 90% Corn Oil
Solubility: ≥ 2.5 mg/mL (3.06 mM); Clear solution

This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown). If the continuous dosing period exceeds half a month, please choose this protocol carefully.
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 900 μL Corn oil, and mix evenly.

In Vivo Dissolution Calculator

Please enter the basic information of animal experiments:

Dosage

mg/kg

Animal weight
(per animal)

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 (sealed storage, away from moisture and 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: 99.82%

Select Batch:

Data Sheet (280 KB) SDS (393 KB)

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

Handling Instructions (2659 KB)

References

[1]. Trinh le A, et al. Fluorescent in situ hybridization employing the conventional NBT/BCIP chromogenic stain. Biotechniques. 2007 Jun;42(6):756-9. [Content Brief]

Complete Stock Solution Preparation Table

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
DMSO / H₂O / 70%DMF	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.

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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Nitro blue tetrazolium chloride (Synonyms: NBT)

Customer Review

Nitro blue tetrazolium chloride Related Antibodies

4 Publications Citing Use of MCE Nitro blue tetrazolium chloride

Featured Recommendations

•Related Small Molecules:

•Related Proteins:

Biological Activity

Purity & Documentation

References

Customer Review

Nitro blue tetrazolium chloride Related Antibodies

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Complete Stock Solution Preparation Table

Nitro blue tetrazolium chloride Related Classifications

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