Shop All Enzyme Substrates

GFP-4E-BP1

4E-BP1 expressed as a fusion with GFP (Green Fluorescent Protein) to act as a physiological substrate for FRAP1 (mTor).

For use in combination with the LanthaScreen® Tb-anti-4E-BP1 [pThr46] Antibody (PV4757) to evaluate inhibitors in medium to high throughput screening applications. An optimized mTor assay buffer (PV4794) is also available.

PIP2:PS Lipid Kinase Substrate

Specifically optimized for use with the Class I PI3 kinases (PIK3CA, PIK3CB, PIK3CD, and PIK3CG), this substrate consists of lipid vesicles of a 1:19 molar ratio of phosphoinositol-4,5-bisphosphate (PIP2) with phosphoserine (PS).

For use with the Adapta™ Universal Kinase Assay (PV5099).

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

This CSPD® 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 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.

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.

PFB-FDGlu (5-(Pentafluorobenzoylamino)Fluorescein Di-β-D-Glucopyranoside) (Invitrogen™)

The ß-glucosidase substrate,

5-(pentafluorobenzoylamino)fluorescein di-ß-D-glucopyranoside (PFB-FDGlu)

yields the green-fluorescent PFB-F dye (excitation/emission maxima ~492/516 nm), which appears to localize to endosomal and lysosomal compartments when loaded into cells by pinocytosis.

DQ™ Green BSA - Special Packaging (Invitrogen™)

DQ Green BSA is a fluorogenic substrate for proteases. A strong fluorescence quenching effect is observed when proteins are heavily labeled with BODIPY dyes. Upon hydrolysis of the DQ Green BSA to single, dye-labeled peptides by proteases, this quenching is relieved, producing brightly fluorescent products.

Pierce™ ECL Western Blotting Substrate (Thermo Scientific™)

Thermo Scientific Pierce ECL Western Blotting Substrate is a value-priced, entry-level horseradish peroxidase (HRP) substrate for enhanced chemiluminescence (ECL) that directly replaces costlier products without the need to re-optimize conditions.

Pierce ECL Plus Western Blotting Substrate characteristics include:
Sensitivity: low picogram
Stability: 1-hr working solution stability, 1-year kit stability at 4°C
Compatibility: nitrocellulose and PVDF membranes
Signal duration: 1 to 2 hours
Recommended primary antibody concentration: 1:1,000–1:5,000 dilution (0.2–10 µg/mL)
Recommended secondary antibody concentration: 1:1,000–1:15,000 dilution (0.07–1.0 µg/mL)

Compare and learn more about our western blot chemiluminescent substrates ›
View all HRP substrates available ›

Pierce ECL Western Blotting Substrate provides reliability and performance equivalent to other standard ECL substrates for detection of HRP enzyme activity. It enables the detection of picogram amounts of antigen and allows for easy detection of HRP using photographic or other imaging methods.

Because the luminol and peroxide reagent formulations are identical to other commercially available substrate products, one can switch to Pierce ECL without needing to optimize probing conditions or incubation protocols. Pierce ECL Western Blotting Substrate provides performance identical to the original Amersham ECL Western Blotting Detection Reagent from GE Healthcare.

Fluorescein-P53 (Ser15) Peptide Substrate

A fluorescein-labeled peptide substrate based upon p53 containing Ser15. p53 is phosphorylated at Ser15 by a wide range of kinases including PI3K/AKT pathway kinases such DNA-PK, ATM, and ATR.

For use in combination with the LanthaScreen® Tb-anti-phospho-p53 [pSer15] antibody (PV5130) to evaluate inhibitors in medium to high throughput screening applications.

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

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.

Alexa Fluor™ 647-Poly-GT Peptide Substrate

A general AlexaFluor647-labeled peptide substrate consisting of a polymer of glutamate, and tyrosine that is used to monitor tyrosine kinase acitivty.

For use in combination with the LanthaScreen® Eu-PY20 Antibody (PV5696) to evaluate inhibitors in medium to high throughput screening applications.

DQ™ Red BSA  - Special Packaging (Invitrogen™)

DQ Red BSA is a fluorogenic substrate for proteases. A strong fluorescence quenching effect is observed when proteins are heavily labeled with BODIPY dyes. Upon hydrolysis of the DQ Red BSA to single, dye-labeled peptides by proteases, this quenching is relieved, producing brightly fluorescent products.

Fluorescein Di-β-D-Glucopyranoside (FDGlu) (Invitrogen™)

Fluorescein di-β-D-glucopyranoside is one of the most sensitive substrates for glucosidases. The nonfluorescent substrate is sequentially hydrolyzed by β-glucosidase, first to fluorescein monoglucoside and then to highly fluorescent fluorescein. Enzyme-mediated hydrolysis of fluorescein di-β-D-glucopyranoside can be followed by the increase in either absorbance or fluorescence.

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.

ONPG (o-nitrophenyl-β-D-galactopyranoside) (Thermo Scientific™)

Thermo Scientific Pierce ONPG is the preferred colorimetric substrate for ELISA applications involving beta-galactosidase (b-Gal) as the reporter enzyme.

When using beta-Galactosidase as the label for proteins in ELISA studies, a wide variety of substrates are available, including: o-Nitrophenyl-beta-D-galactopyranosidase (ONPG), Naphthol-AS-Bl-beta-D-galactopyranosidase (Nap-Gal), and 4-Methyl-umbelliferyl-beta-D-galactopyranosidase (MUm-Gal). However, it is important to choose a substrate with adequate solubility, that uses readily available equipment, and that gives a significant reading over the background. ONPG is a superior beta-Galactosidase substrate option.(1) The product formed is completely soluble and has a high extinction coefficient at 405nm. The substrate yields a yellow product that is easily detectable in the visual range after stopping the reaction with 1M Sodium Carbonate.