Shop All Alkaline Phosphatase Substrates

1-Step™ NBT/BCIP plus Suppressor Substrate Solution (Thermo Scientific™)

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 Substrate Solution
NBT Substrate Powder (nitro-blue tetrazolium chloride)
BCIP Substrate Powder (5-bromo-4-chloro-3'-indolyphosphate p-toluidine)

ELISA-Light™ Immunoassay System with CSPD™ and Emerald-II™ Substrate/Enhancer Solution (Applied Biosystems™)

The ELISA-Light™ Immunoassay System with CSPD® and Emerald-II™ Substrate⁄Enhancer Solution is used for ultra-sensitive detection of antigens or antibodies in multiple immunoassay formats employing an alkaline phosphatase enzyme label.

• Heterogeneous assay format increases sensitivity and dynamic range and reduces interference by test compounds in the detection portion of the assay (as compared with homogeneous assay formats)
• Chemiluminescent immunoassay detection allows greater sensitivity and improved dynamic range as compared to radioimmunoassays and immunoassays that employ colorimetric or fluorescent detection
• High sensitivity and wide dynamic range permit sensitive detection with fewer assays, because sample dilution to fit the dynamic range of the assay is unnecessary
• A variety of ready-to-use substrate formulations lets the user precisely optimize signal intensity to the exact requirements of an assay
• The kinetics of the glow-assay provides a "window" during which measurements may be performed, facilitating HTS applications where assay automation is used
• Chemiluminescent immunoassay detection provides versatile applications such as sandwich immunoassays, competitive immunoassays, and DNA probe capture assays
• Non-radioactive immunoassays eliminate concerns over use of radioisotopes

Extend the Limits of Detection
The ELISA-Light™ system offers an excellent combination of high sensitivity and throughput for assays measured in a luminometer. The ELISA-Light™ system significantly extends the limits of detection of most immunoassays previously performed with colorimetric and isotopic methods.

Multiple Substrate⁄Enhancer Formulations
The kit contains reagents optimized for chemiluminescent ELISAs employing alkaline phosphatase labels and is available with a choice of ready-to-use chemiluminescent substrate formulations containing CSPD® or CDP-Star® substrate with Sapphire-II™ or Emerald-II™ luminescence enhancers. Multiple substrate⁄enhancer formulations provide options that let the user optimize specific assays or luminometers. The specific formulation that will provide the highest sensitivity for a specific assay can vary. Therefore, an ELISA sampler kit containing all four substrate⁄enhancer formulations is available to facilitate assay development.

For Research Use Only. Not for use in diagnostics procedures.

NBT/BCIP Reagent Kit (Invitrogen™)

The NBT/BCIP Reagent Kit includes 1 g samples of both NBT and BCIP for the chromogenic detection of phosphatase.

CDP-Star™ Substrate (0.25 mM Ready-To-Use) (Invitrogen™)

The CDP-Star® Substrate is a chemiluminescent alkaline phosphatase substrate, ready-to-use for protein or nucleic acid blotting on nitrocellulose membranes. This substrate can also be used in solution-based assays.

This CDP-Star® chemiluminescent substrate for alkaline phosphatase let you detect alkaline phosphatase and alkaline phosphatase-labeled molecules with unparalleled sensitivity, speed, and ease.

• Chemiluminescent substrate provides highly sensitive replacements for the widely used fluorogenic substrate methylumbelliferyl phosphate (MUP), and the colorimetric substrate p-nitrophenyl phosphate (pNPP).
• Low background luminescence coupled with high-intensity light output enables detection of alkaline phosphatase labels with the highest possible sensitivity and signal-to-noise.

This versatile chemiluminescent substrate exhibits high sensitivity in membrane-based applications such as Southern, Northern, and Western blotting. This substrate can also be used in solution-based assays such as immunoassays, DNA probe assays, enzyme assays, and reporter gene assays. Maximum light levels are reached in approximately 10 minutes and glow emission persists for several hours.

For Research Use Only. Not for use in diagnostics procedures.

pNPP Powder (Invitrogen™)

pNPP may be used as a substrate for alkaline phosphatase (AP) in enzyme immunoassays.

CSPD™ Substrate (25 mM Concentrate) (Invitrogen™)

This concentrated CSPD® substrate is a chemiluminescent alkaline phosphatase substrate for protein or nucleic acid blotting on nitrocellulose membranes. This substrate can also be used in solution-based assays.

This CSPD® chemiluminescent substrate for alkaline phosphatase let you detect alkaline phosphatase and alkaline phosphatase-labeled molecules with unparalleled sensitivity, speed, and ease.

• Chemiluminescent substrates provide highly sensitive replacements for the widely used fluorogenic substrate methylumbelliferyl phosphate (MUP), and the colorimetric substrate p-nitrophenyl phosphate (pNPP).
• Low background luminescence coupled with high-intensity light output enables detection of alkaline phosphatase labels with the highest possible sensitivity and signal-to-noise.

This versatile chemiluminescent substrate exhibits high sensitivity in membrane-based applications such as Southern, Northern, and Western blotting. This substrate can also be used in solution-based assays such as immunoassays, DNA probe assays, enzyme assays, and reporter gene assays. Maximum light levels are reached in approximately 10 minutes and glow emission persists for several hours. Solution assays require chemiluminescent enhancers. Membrane-based assays may also require them for increased light output and sensitivity.

For Research Use Only. Not for use in diagnostics procedures.

CDP-Star™ Substrate (12.5 mM Concentrate) (Invitrogen™)

This concentrated CDP-Star® Substrate is a chemiluminescent alkaline phosphatase substrate for protein or nucleic acid blotting on nitrocellulose membranes. This substrate can also be used in solution-based assays.

This CDP-Star® chemiluminescent substrate for alkaline phosphatase let you detect alkaline phosphatase and alkaline phosphatase-labeled molecules with unparalleled sensitivity, speed, and ease.

• Chemiluminescent substrate provides highly sensitive replacements for the widely used fluorogenic substrate methylumbelliferyl phosphate (MUP), and the colorimetric substrate p-nitrophenyl phosphate (pNPP).
• Low background luminescence coupled with high-intensity light output enables detection of alkaline phosphatase labels with the highest possible sensitivity and signal-to-noise.

This versatile chemiluminescent substrate exhibits high sensitivity in membrane-based applications such as Southern, Northern, and Western blotting. This substrate can also be used in solution-based assays such as immunoassays, DNA probe assays, enzyme assays, and reporter gene assays. Maximum light levels are reached in approximately 10 minutes and glow emission persists for several hours.

For Research Use Only. Not for use in diagnostics procedures.

DynaLight™ Trigger Solution (Invitrogen™)

Achieve faster ELISA results with DynaLight™ Trigger Solution when used in combination with DynaLight™ Substrate with RapidGlow™ Enhancer, a ready-to-use chemiluminescent substrate formulation that has been optimized to achieve faster results in solution-based assays.

Features of DynaLight™ Substrate with RapidGlow™ Enhancer:

• Faster kinetics and higher signal levels than colorimetric, fluorescent, or other chemiluminescent chemistries
• Sensitive attomole detection of purified alkaline phosphatase enzyme
• Optimized formulation for both microplate and bead-based assays
• Substrate stability of over 1 year at 4°C
• Compatible with automated platforms with trigger solution/positions already in place

The DynaLight™ Substrate with RapidGlow™ Enhancer provides a robust chemiluminescent reaction when catalyzed by alkaline phosphatase. This reaction achieves maximum light emission within 2 minutes and maintains a sustained signal for hours. When combined with DynaLight™ Trigger Solution, DynaLight™ Substrate with RapidGlow™ Enhancer delivers maximum signal within seconds. The environmentally-friendly DynaLight™ Trigger Solution provides maximum signal within 1 second of addition, with a longer half-life to its ground state than other chemiluminescent Flash technologies, allowing ample time to collect data reliably.

For Research Use Only. Not for human or animal therapeutic or diagnostic use.

ELISA-Light™ Immunoassay System with CDP-Star™ and Sapphire-II™ Substrate/Enhancer Solution (Applied Biosystems™)

The ELISA-Light™ Immunoassay System with CDP-Star® and Sapphire-II™ Substrate⁄Enhancer Solution is used for ultra-sensitive detection of antigens or antibodies in multiple immunoassay formats employing an alkaline phosphatase enzyme label.

• Heterogeneous assay format increases sensitivity and dynamic range and reduces interference by test compounds in the detection portion of the assay (as compared with homogeneous assay formats)
• Chemiluminescent immunoassay detection allows greater sensitivity and improved dynamic range as compared to radioimmunoassays and immunoassays that employ colorimetric or fluorescent detection
• High sensitivity and wide dynamic range permit sensitive detection with fewer assays, because sample dilution to fit the dynamic range of the assay is unnecessary
• A variety of ready-to-use substrate formulations lets the user precisely optimize signal intensity to the exact requirements of an assay
• The kinetics of the glow-assay provides a "window" during which measurements may be performed, facilitating HTS applications where assay automation is used
• Chemiluminescent immunoassay detection provides versatile applications such as sandwich immunoassays, competitive immunoassays, and DNA probe capture assays
• Non-radioactive immunoassays eliminate concerns over use of radioisotopes

Extend the Limits of Detection
The ELISA-Light™ system offers an excellent combination of high sensitivity and throughput for assays measured in a luminometer. The ELISA-Light™ system significantly extends the limits of detection of most immunoassays previously performed with colorimetric and isotopic methods.

Multiple Substrate⁄Enhancer Formulations
The kit contains reagents optimized for chemiluminescent ELISAs employing alkaline phosphatase labels and is available with a choice of ready-to-use chemiluminescent substrate formulations containing CSPD® or CDP-Star® substrate with Sapphire-II™ or Emerald-II™ luminescence enhancers. Multiple substrate⁄enhancer formulations provide options that let the user optimize specific assays or luminometers. The specific formulation that will provide the highest sensitivity for a specific assay can vary. Therefore, an ELISA sampler kit containing all four substrate⁄enhancer formulations is available to facilitate assay development.

For Research Use Only. Not for use in diagnostics procedures.

1-Step™ PNPP Substrate Solution (Thermo Scientific™)

Thermo Scientific Pierce PNPP (p-nitrophenyl phosphate) is a colorimetric, soluble substrate of alkaline phosphatase for use in ELISA applications. This product is a ready-to-use (1X) proprietary buffered solution of PNPP.

Features of PNPP (p-nitrophenyl phosphate):

• One-component alkaline phosphatase (AP) substrate
• Absorbance maximum of 405nm
• Develops a yellow hue in solution upon reaction with AP
• Safer to use—contains a non-mercury, non-azide preservative

Until now, the method for preparing PNPP solutions required dissolving the powder or tablets in buffer and then diluting the solution to the desired concentration. Substrate instability made it necessary to prepare it on a daily basis. Pierce 1-Step PNPP circumvents this time-consuming and variable introducing step by providing a single component PNPP substrate. This substrate is stable for 12 months at 2°C-8°C and can be stopped with conventional methods.

PNPP (p-Nitrophenyl Phosphate, Disodium Salt) is a widely used substrate for detecting alkaline phosphatase in ELISA applications. When alkaline phosphatase and PNPP are reacted, a yellow water-soluble reaction product is formed. This reaction product absorbs light at 405nm. The substrate is offered in several formats.

Related Products
Pierce™ PNPP Substrate Kit
PNPP Substrate Tablets
PNPP Substrate

Luminescence Enhancer For Solution Assays, Emerald™ Enhancer

The Emerald™ chemiluminescence enhancer
is a concentrated reagent suitable for use with chemiluminescent substrates (AMPPD®, CSPD®, Galacton®, Glucon™, or Glucuron®).

For Research Use Only. Not for use in diagnostics procedures.

CDP-Star™ Substrate (0.4 mM Ready-To-Use) with Sapphire-II™ Enhancer, Size A (Invitrogen™)

The CDP-Star® Substrate with Sapphire-II™ Enhancer is a chemiluminescent alkaline phosphatase substrate, ready-to-use for protein or nucleic acid blotting on nitrocellulose membranes. This substrate can also be used in solution-based assays.

This CDP-Star® chemiluminescent substrate for alkaline phosphatase let you detect alkaline phosphatase and alkaline phosphatase-labeled molecules with unparalleled sensitivity, speed, and ease.

• Chemiluminescent substrate provides highly sensitive replacements for the widely used fluorogenic substrate methylumbelliferyl phosphate (MUP), and the colorimetric substrate p-nitrophenyl phosphate (pNPP).
• Low background luminescence coupled with high-intensity light output enables detection of alkaline phosphatase labels with the highest possible sensitivity and signal-to-noise.

This versatile chemiluminescent substrate exhibits high sensitivity in membrane-based applications such as Southern, Northern, and Western blotting. This substrate can also be used in solution-based assays such as immunoassays, DNA probe assays, enzyme assays, and reporter gene assays. Maximum light levels are reached in approximately 10 minutes and glow emission persists for several hours. Solution assays require chemiluminescent enhancer. Membrane-based assays may also require it for increased light output and sensitivity.

For Research Use Only. Not for use in diagnostics procedures.

pNPP Solution (Invitrogen™)

pNPP may be used as a substrate for alkaline phosphatase (AP) in enzyme immunoassays.

CSPD™ Substrate (0.25 mM Ready-To-Use) with Nitro-Block™ Enhancer (Invitrogen™)

This CSPD® chemiluminescent substrate with Nitro-Block™ Enhancer is a chemiluminescent alkaline phosphatase substrate, ready-to-use for protein or nucleic acid blotting on nitrocellulose membranes. This substrate can also be used in solution-based assays.

This CSPD® chemiluminescent substrate for alkaline phosphatase let you detect alkaline phosphatase and alkaline phosphatase-labeled molecules with unparalleled sensitivity, speed, and ease.

• Chemiluminescent substrates provide highly sensitive replacements for the widely used fluorogenic substrate methylumbelliferyl phosphate (MUP), and the colorimetric substrate p-nitrophenyl phosphate (pNPP).
• Low background luminescence coupled with high-intensity light output enables detection of alkaline phosphatase labels with the highest possible sensitivity and signal-to-noise.

This versatile chemiluminescent substrate exhibits high sensitivity in membrane-based applications such as Southern, Northern, and Western blotting. This substrate can also be used in solution-based assays such as immunoassays, DNA probe assays, enzyme assays, and reporter gene assays. Maximum light levels are reached in approximately 10 minutes and glow emission persists for several hours. Solution assays require chemiluminescent enhancers. Membrane-based assays may also require them for increased light output and sensitivity. Solution assays require chemiluminescent enhancer. Membrane-based assays may also require it for increased light output and sensitivity.

For Research Use Only. Not for use in diagnostics procedures.

DynaLight™ Substrate with RapidGlow™ Enhancer (Invitrogen™)

Achieve faster ELISA results with DynaLight™ Substrate with RapidGlow™ Enhancer, a ready-to-use chemiluminescent substrate formulation that has been optimized to achieve faster results in solution-based assays. The DynaLight™ Substrate with RapidGlow™ Enhancer formulation includes 1,2-dioxetane chemiluminescent substrate and a polymeric enhancer that enables ultra-sensitive immunoassay detection by alkaline phosphatase label.

Features of DynaLight™ Substrate with RapidGlow™ Enhancer:

• Faster kinetics and higher signal levels than colorimetric, fluorescent, or other chemiluminescent chemistries
• Sensitive attomole detection of purified alkaline phosphatase enzyme
• Optimized formulation for both microplate and bead-based assays
• Substrate stability of over 1 year at 4°C
• Compatibility with automated platforms with trigger solution/positions already in place

The DynaLight™ Substrate with RapidGlow™ Enhancer provides a robust chemiluminescent reaction when catalyzed by alkaline phosphatase. This reaction achieves maximum light emission within 2 minutes and maintains a sustained signal for hours. When combined with DynaLight™ Trigger Solution, DynaLight™ Substrate with RapidGlow™ Enhancer delivers maximum signal within seconds. The environmentally-friendly DynaLight™ Trigger Solution provides maximum signal within 1 second of addition, with a longer half-life to its ground state than other chemiluminescent Flash technologies, allowing ample time to collect data reliably.

For Research Use Only. Not for human or animal therapeutic or diagnostic use.