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Roche

NBT

4-Nitro blue tetrazolium chloride, solution

Synonym(s):

Nitroblue tetrazolium chloride

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About This Item

UNSPSC Code:
41116100

Quality Level

100

Assay

>95%

form

solution

mol wt

Mr 817.7

packaging

pkg of 3 mL (300 mg)

manufacturer/tradename

Roche

technique(s)

Northern blotting: suitable
Southern blotting: suitable
hybridization: suitable

storage temp.

−20°C

General description

NBT is a chromogenic and soluble substance. It is a substrate of alkaline phosphatase (AP) and is converted to an indigo/purple dye on its action.

Application

  • Immunohistocytochemistry
  • Western blot
  • Southern blot
  • Northern blot
  • Colony and plaque hybridization
  • In situ hybridization

Physical form

Solution (100 mg/ml) in 70% dimethylformamide (DMF) (v/v). The color of the NBT solution can vary between yellow and brown depending on the lot used.

Note: The color does not impair the quality or the function of the reagent. In some cases a precipitate may occur which is easily brought into solution by briefly warming the substrate at 37 °C.

Preparation Note

Working solution: Preparation of staining solution
  • Always prepare this solution fresh shortly before use!For all applications except DIG System: Add 50 μl NBT solution and 37,5 μl BCIP to 10 ml 0.1 M Tris-HCl, pH 9.5 (20 °C), 0.1 M NaCl, 0.05 M MgCl2.
  • For DIG System applications: Add 50 μl NBT solution and 37,5 μl BCIP to 10 ml 0.1 M Tris-HCl, pH 9.5 (20 °C), 0.1 M NaCl
  • Note: Do not include MgCl2 in the DIG detection buffer as this might lead to spotty background on the membrane after the detection procedure. Alkaline Phosphatase does not require Mg2+.

Other Notes

For life science research only. Not for use in diagnostic procedures.

Pictograms

Exclamation markHealth hazard
GHS07,GHS08

Signal Word

Danger

Hazard Statements

H319,H332,H360D

Hazard Classifications

Acute Tox. 4 Inhalation - Eye Irrit. 2 - Repr. 1B

Storage Class Code

6.1C - Combustible acute toxic Cat.3 / toxic compounds or compounds which causing chronic effects

WGK

WGK 2

Flash Point(F)

167.0 °F

Flash Point(C)

75 °C

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Protocols

NBT Protocol & Troubleshooting

There are several counterstains possible in combination with BM Purple (or NBT/BCIP in general), including FastGreen FCF and Nuclear Fast Red.

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

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