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1-Step™ NBT/BCIP Substrate Solution
1-Step™ NBT/BCIP Substrate Solution
Thermo Scientific™

1-Step™ NBT/BCIP Substrate Solution

Thermo Scientific Pierce NBT and BCIP Substrates are powders and ready-made solutions of nitro-blue tetrazolium and 5-bromo-4-chloro-3'-indolyphosphate for chromogenic blotRead more
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Catalog NumberQuantity
34042250 mL
Catalog number 34042
Price (USD)
218.00
Each
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250 mL
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Price (USD)
218.00
Each
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Thermo Scientific Pierce NBT and BCIP Substrates are powders and ready-made solutions of nitro-blue tetrazolium and 5-bromo-4-chloro-3'-indolyphosphate for chromogenic blot and IHC staining with alkaline phosphatase probes.

The combination of NBT (nitro-blue tetrazolium chloride) and BCIP (5-bromo-4-chloro-3'-indolyphosphate p-toluidine salt) yields an intense, insoluble black-purple precipitate when reacted with alkaline phosphatase, a popular enzyme conjugate for antibody probes. Pierce NBT and BCIP powders are available individually and in two convenient, ready-to-use solutions (with and without a levamisole suppressor of endogenous phosphatase activity).

Features:

AP substrate—for detection of alkaline phosphatase (alk-phos) activity on solid media, including nitrocellulose and PVDF membranes and fixed tissue samples
Chromogenic—no special equipment needed for visualization; produces purple-black precipitate that is easy to photograph
Package options—choose individual NBT and BCIP powders, or preformulated, ready-to-use solutions with or without levamisole suppressor

Highlights of Pierce NBT/BCIP 1-Step Solution (Part No. 34042):
• Best for immunoblotting applications
• Ready-to-use single component
• Sensitive with low background and minimial assay-to-assay variability

Highlights of Pierce NBT/BCIP plus Suppressor 1-Step Solution (Part No. 34070):
• Best for immunohistochemistry applications
• Ready-to-use single component
• Contains 1 mM levamisole to inhibit endogenous phosphatase activity, while yielding an intense dark purple precipitate in the presence of calf intestinal phosphatase

Related Products
1-Step™ NBT/BCIP plus Suppressor Substrate Solution
NBT Substrate Powder (nitro-blue tetrazolium chloride)
BCIP Substrate Powder (5-bromo-4-chloro-3'-indolyphosphate p-toluidine)
For Research Use Only. Not for use in diagnostic procedures.
Specifications
DescriptionPierce 1-Step NBT/BCIP Substrate Solution
ReactivityAlkaline Phosphatase
Detection MethodColorimetric
FormLiquid
Membrane CompatibilityNitrocellulose, PVDF
Product Line1-Step™
Product TypeSubstrate
Quantity250 mL
SubstrateAlkaline Phosphatase
Unit SizeEach
Contents & Storage
Upon receipt store product at 4°C. Product is shipped at ambient temperature.
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Certificates

Lot #Certificate TypeDateCatalog Number(s)
3214583Certificate of AnalysisMay 19, 202534042
AC409312Certificate of AnalysisApr 11, 202534042
AA407986Certificate of AnalysisFeb 24, 202534042
AA404446Certificate of AnalysisFeb 03, 202534042-100UG, 34042
ZJ403417Certificate of AnalysisDec 16, 202434042
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Safety Data Sheets

Frequently asked questions (FAQs)

For colorimetric alkaline phosphatase substrates, we offer the following products:

Novex AP Chromogenic Substrate (BCIP/ NBT) (Cat. No. WP20001)

1-Step NBT/BCIP Substrate Solution (Cat. No. 34042)

1-Step PNPP Substrates Solution (Cat. No. 37621)

NBT and BCIP are also available as standalone products:

NBT Substrate Powder (nitro-blue tetrazolium chloride) (Cat. No. 34035)

BCIP Substrate Powder (5-bromo-4-chloro-3'-indolylphosphate p-toluidine) (Cat. No. 34040)

For chemiluminescent alkaline substrates, we offer the following products:

Novex AP Chemiluminescent Substrate (CDP-Star) (Cat. No. WP20002)

NovaBright Phospha-Light EXP Assay Kit for SEAP (Secreted Placental Alkaline Phosphatase) Reporter Gene Detection (Cat. Nos. N10577, N10578)

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Yes, it will work with both PVDF as well as nitrocellulose membranes

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

To eliminate endogenous phosphatase activity, add a final concentration of 1 mM levamisole to the 1-Step NBT/BCIP Substrate Solution.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

No, the 1-Step NBT/BCIP Substrate Solution cannot be used for ELISA. It is a precipitating alkaline phoshpatase substrate developed for immunoblotting, immunohistochemistry and in situ hybridzation. For ELISA, use our 1-Step PNPP Substrate Solution (Cat. No. 37621).

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

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