Shop All Alkaline Phosphatase Substrates

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 (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.

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.

Luminescence Enhancer For Solution Assays, Sapphire™ Enhancer (Invitrogen™)

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

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

CSPD™ Substrate (0.25 mM Ready-To-Use) (Invitrogen™)

This CSPD® 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 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.

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

Novex™ AP Chromogenic Substrate (BCIP⁄ NBT) (Invitrogen™)

Novex® AP Chromogenic Substrate is a ready to use single component reagent for sensitive immunodetection of alkaline phosphatase (AP) enzyme on western blot and dot blot membranes. Compatable for use with Western Breeze™ protocols, this ready-to-use solution of 5-bromo-4-chloro-3-indolyl-1-phosphate (BCIP) and nitroblue tetrazolium (NBT) will forms a black-purple precipitate upon reaction with alkaline phosphatase.

1-Step™ NBT/BCIP 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 plus Suppressor Substrate Solution
NBT Substrate Powder (nitro-blue tetrazolium chloride)
BCIP Substrate Powder (5-bromo-4-chloro-3'-indolyphosphate p-toluidine)

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.

Novex™ AP Chemiluminescent Substrate (CDP-Star™) (Invitrogen™)

Novex® AP Chemiluminescent Substrate is a non-radioactive ready-to-use solution of CDP-Star for chemiluminescence-based immunodetection of alkaline phosphatase (AP) on western blot or dot blot membranes. This reagent provides detection sensitivities superior to that of precipitating chromogenic substrates. Low picogram levels of detection can be achieved using either X-ray film or imaging equipment.

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

CSPD™ Substrate (0.4 mM Ready-To-Use) with Emerald-II™ Enhancer, Size A (Invitrogen™)

This CSPD® substrate with Emerald-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 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.

PNPP Substrate (Thermo Scientific™)

Thermo Scientific Pierce PNPP (p-nitrophenyl phosphate) is a colorimetric, soluble substrate of alkaline phosphatase for use in ELISA applications.

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

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
1-Step™ PNPP Substrate Solution
Pierce™ PNPP Substrate Kit
PNPP Substrate Tablets

Luminescence Enhancer For Membrane Blotting Assays, Nitro-Block-II™ Enhancer (Applied Biosystems™)

This concentrated Nitro-Block-II™ Enhancer can be used with CSPD® or CDP-Star® chemiluminescent substrates to increase the alkaline phosphatase signal for protein or nucleic acid blots.

The Nitro-Block-II™ membrane enhancer increases chemiluminescent signal intensity on nitrocellulose and PVDF membranes.

• Chemically optimized formulations provide enhanced signal intensity in blotting applications with dioxetane-based substrate solutions (CSPD® or CDP-Star® substrates) on nitrocellulose or PVDF membranes.
• Enhancer increases signal intensity, allowing decreased signal acquisition time.

Increased Signal, Shorter Exposures
Without enhancer, the chemiluminescent signal on nitrocellulose membranes is very weak, making detection either impossible or very time consuming. With enhancer, short exposures (10 to 45 minutes) can be performed on the same material. Although PVDF membranes do not require the use of Nitro-Block-II™ enhancer, exposure times are ten-fold faster with enhancer. This permits very short exposure times on PVDF, ranging from 15 seconds to 15 minutes for western blots. Nitro-Block-II™ enhancer may be used with CSPD® substrate on nitrocellulose or PVDF membranes. Nitro-Block-II™ enhancer should only be used with CDP-Star® substrate on nitrocellulose membranes.

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

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.

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

The CDP-Star® Substrate with Emerald-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.