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X-Gluc (Thermo Scientific™)

Thermo Scientific X-Gluc (5-bromo-4-chloro-3-indolyl-beta-D-glucuronic acid, cyclohexylammonium salt) is a substrate for beta-glucuronidase (GUS) which is encoded by gusA, a widely used reporter gene. Glucuronidase cleaves X-Gluc to produce colorless glucuronic acid and an intense blue precipitate of chloro-bromoindigo.

Applications

• Detection of GUS expression in plant cells and tissues
• Detection of infections caused by E. coli
• Detection of bacterial contamination in food and water samples.

Note
• Preparation of a 0.1 M stock solution in dimethylformamide or dimethylsulfoxide is recommended.

Vivid™ BOMR Substrate

Vivid® Substrates are blocked dyes that yield a minimal fluorescence signal until oxidation occurs. Cytochrome P450 metabolism at either of two potential oxidation sites releases the highly fluorescent product. They have superior fluorescence, solubility, and kinetic properties compared to conventional fluorogenic probes. This results in higher sensitivity, greater signal-to-noise ratio, and better assay reproducibility. When used in combination with CYP450 BACULOSOMES® Plus Reagents as part of the Vivid® CYP450 Screening Kits, the Vivid® Substrates enable rapid, robust detection of cytochrome P450 inhibitors. This particular substrate (Vivid ® BOMR Substrate) yields a product that emits a red fluorescence.

More About Vivid® CYP450 Screening Kits
Vivid® CYP450 Screening Kits are high-throughput fluorescence-based assays for detection of enzyme-drug interactions and CYP450 inhibition. Vivid® CYP450 Screening Kits include Vivid® Substrate, Vivid® Fluorescent Standard, reaction buffer, Vivid® Regeneration System, NADP+, and CYP450 BACULOSOMES® Plus Reagent. CYP450 BACULOSOMES® Plus Reagents are microsomes prepared from insect cells expressing a human CYP450 isozyme and human cytochrome P450 reductase. CYP450 BACULOSOMES® Plus Reagents offer a distinct advantage over human liver microsomes in that only one CYP450 isozyme is expressed, thereby preventing metabolism by other CYP450s.

>> View all Vivid® CYP450 Screening Kits

Applications: high-throughput screening of enzyme-drug interactions, compound profiling for drug inhibition of cytochrome P450 isozymes, generation of predictive SAR models to guide compound acquisition

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

X-Gal Solution, ready-to-use (Thermo Scientific™)

Thermo Scientific X-Gal (5-bromo-4-chloro-3-indolyl-beta-D-galacto-pyranoside) is an inert chromogenic substrate for beta-galactosidase which hydrolyzes X-Gal into colorless galactose and 4-chloro-3-brom-indigo, forming an intense blue precipitate. Induction of the lacZ gene with IPTG leads to the hydrolysis of X-Gal and to the development of blue colonies (see Supporting data).

X-Gal Solution, ready-to-use, is stable, 0.22 µm membrane filtered solution formulated for direct use in conjunction with IPTG for blue/white colony screening.

Applications

• Blue/white colony screening to distinguish recombinant (white) from non-recombinant (blue) colonies
• Visualization of beta-galactosidase reporter gene expression in transfected eukaryotic cells
• Detection of beta-galactosidase activity in immunological and histochemical procedures

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X-Gal

CCF2-AM

CCF2-AM, our LiveBLAzer™ substrate, is a lipophilic, esterified form of the CCF2 substrate which allows it to readily enter cells. Upon entry, cleavage by endogenous cytoplasmic esterases rapidly converts CCF2-AM into its negatively charged form, CCF2 or CCF4, which is retained in the cytosol. CCF2 is a Fluorescence Resonance Energy Transfer (FRET) substrate which consists of a cephalosporin core linking 7-hydroxycoumarin to fluorescein. This product is to be used as a FRET-based substrate for β-lactamase as a sensitive reporter of mammalian gene expression in the GeneBLAzer® technology.

SuperSignal™ West Atto Ultimate Sensitivity Substrate (Thermo Scientific™)

SuperSignal West Atto Ultimate Sensitivity Chemiluminescent Substrate is an ultra-sensitive enhanced chemiluminescent (ECL) substrate for high attogram protein detection. SuperSignal West Atto substrate provides the highest-level sensitivity and better signal-to–noise ratios compared to other commercially available high-performance horseradish peroxidase (HRP) substrates.

SuperSignal West Atto Ultimate Sensitivity Chemiluminescent Substrate characteristics include:
• Sensitivity: low-femtogram to high-attogram
• Stability: eight hours for working solution, one year for kit at 4°C
• Compatibility: nitrocellulose and PVDF membranes
• Signal duration: up to four hours
• Recommended primary antibody concentration: 1:1,000–1:5,000 dilution (0.2–1.0 µg/mL)
• Recommended secondary antibody concentration: 1:100,000–1:250,000 dilution (4–10 ng/mL)

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

SuperSignal West Atto Ultimate Sensitivity Chemiluminescent Substrate is designed for superior sensitivity and signal-to–noise ratios for very low abundant targets or precious samples in western blot analysis. When combined with optimized antibody concentrations and blocking buffers, SuperSignal West Atto substrate enables detection of target proteins in amounts that are too small to be seen with typical high performance ECL substrates.

AEC Solution (IHC)

This Single Solution utilizes aminoethyl carbazole (AEC), an alcohol soluble chromogen, and generates a reddish deposit at the reaction site on tissue or cell preparations. Single Solution AEC is suitable for use with manual or automated staining systems, and is provided as a liquid in a single bottle that can be stored at -8°C.

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.

Amplex™ UltraRed Reagent (Invitrogen™)

Our Amplex UltraRed reagent improves upon the performance of our unique Amplex Red reagent, offering brighter fluorescence and enhanced sensitivity on a per-mole basis in peroxidase or peroxidase-coupled enzyme assays. The Amplex UltraRed reagent is also less sensitive to pH, and exhibits improved stability in the presence of hydrogen peroxide (H2O2) or thiols such as dithiothreitol (DTT).

Fluorescein Di-β-D-Glucuronide (FDGlcU) (Invitrogen™)

Fluorescein di-β-D-glucuronide (FDGlcU) is colorless and nonfluorescent until it is hydrlozed to the monoglucuronide and then to the highly fluorescent fluorescein.

Vivid™ OOMR Substrate

Vivid® Substrates are blocked dyes that yield a minimal fluorescence signal until oxidation occurs. Cytochrome P450 metabolism at either of two potential oxidation sites releases the highly fluorescent product. They have superior fluorescence, solubility, and kinetic properties compared to conventional fluorogenic probes. This results in higher sensitivity, greater signal-to-noise ratio, and better assay reproducibility. When used in combination with CYP450 BACULOSOMES® Plus Reagents as part of the Vivid® CYP450 Screening Kits, the Vivid® Substrates enable rapid, robust detection of cytochrome P450 inhibitors. This particular substrate (Vivid ® OOMR Substrate) yields a product that emits a red fluorescence.

More About Vivid® CYP450 Screening Kits
Vivid® CYP450 Screening Kits are high-throughput fluorescence-based assays for detection of enzyme-drug interactions and CYP450 inhibition. Vivid® CYP450 Screening Kits include Vivid® Substrate, Vivid® Fluorescent Standard, reaction buffer, Vivid® Regeneration System, NADP+, and CYP450 BACULOSOMES® Plus Reagent. CYP450 BACULOSOMES® Plus Reagents are microsomes prepared from insect cells expressing a human CYP450 isozyme and human cytochrome P450 reductase. CYP450 BACULOSOMES® Plus Reagents offer a distinct advantage over human liver microsomes in that only one CYP450 isozyme is expressed, thereby preventing metabolism by other CYP450s.

>> View all Vivid® CYP450 Screening Kits

Applications: high-throughput screening of enzyme-drug interactions, compound profiling for drug inhibition of cytochrome P450 isozymes, generation of predictive SAR models to guide compound acquisition

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

QuantaBlu™ NS/K Fluorogenic Substrate Kit (Thermo Scientific™)

The Thermo Scientific QuantaBlu NS/K Fluorogenic Substrate Kit is a two-component kit containing proprietary fluorescent substrate and peroxide solutions for non-stopped kinetic assays. QuantaBlu Fluorogenic Peroxidase Substrates allow rapid kinetic ELISA assays to be performed with greater sensitivity than TMB and OPD substrates

Features of the QuantaBlu NS/K Fluorogenic Substrate Kit:

• More sensitive than TMB, OPD or ABTS substrates
• Large dynamic range (4 log peroxidase concentration range)
• Excellent stability: working solution is stable for 24 hours
• Large stokes shift; excitation/emission maxima of 325/420; range of 315-345/370-460
• Does not photobleach

A variety of substrates are available for detecting peroxidase activity in ELISA-based assays. Colorimetric substrates (e.g., TMB, OPD and ABTS) have been widely used for years. Each of these substrates varies greatly with respect to its performance characteristics such as detection sensitivity, working range and attainable signal-to-noise ratios.

QuantaBlu Fluorogenic Peroxidase Substrate meets all the requirements of an ideal substrate for use in peroxidase detection. QuantaBlu Substrate generates a blue fluorescent product upon reaction with peroxidase. The fluorescent product does not photobleach. Fluorometric-based detection overcomes the limitations of colorimetric substrate detection, which does not allow for quantitation of greater than four optical density units. QuantaBlu Substrate exhibits a flat baseline in assays, which facilitates low-level detection sensitivity and allows for high signal-to-noise ratios.

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QuantaBlu™ Fluorogenic Peroxidase Substrate Kit

X-Gal (Thermo Scientific™)

Thermo Scientific X-Gal (5-bromo-4-chloro-3-indolyl-beta-D-galacto-pyranoside) is an inert chromogenic substrate for beta-galactosidase which hydrolyzes X-Gal into colorless galactose and 4-chloro-3-brom-indigo, forming an intense blue precipitate. Induction of the lacZ gene with IPTG leads to the hydrolysis of X-Gal and to the development of blue colonies (see Supporting data).

Applications

• Blue/white colony screening to distinguish recombinant (white) from non-recombinant (blue) colonies (see Reference 1)
• Visualization of beta-galactosidase reporter gene expression in transfected eukaryotic cells
• Detection of beta-galactosidase activity in immunological and histochemical procedures

Note
• Preparation of a 20 mg/mL stock solution in dimethylformamide (DMF) or dimethylsulfoxide (DMSO) is recommended for X-Gal. DMF dissolves some plastic materials. The direct addition of DMF containing solution to plastic Petri dishes should be avoided.
• DMF is toxic. Allow X-Gal solution containing dimethylformamide to completely dry before plating bacteria on agar plates.

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X-Gal
X-Gal Solution, ready-to-use

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.

LiveBLAzer™ FRET-B/G Loading Kit with CCF2-AM

Background:
GeneBLAzer® cell-based assays utilize the membrane-permeant ester forms (CCF2-AM and CCF4-AM) of the negatively charged fluorescent beta-lactamase substrates, CCF2 and CCF4. These lipophilic esters readily enter the cell, where cleavage by endogeneous cytoplasmic esterases rapidly converts them into their negatively charged forms, thereby trapping them in the cytosol.

Detection of GeneBLAzer® assays is FRET-based. Each substrate is labeled with two fluorophores that form an efficient FRET pair. In the absence of beta-lactamase activity, exciting the coumarin at 409 nm in the intact CCF2 molecule results in FRET to the fluorescein, which emits a green fluorescence signal at 518 nm (Figure 1). In the presence of beta-lactamase activity, however, cleavage of CCF2 spatially separates the two dyes and disrupts FRET, so that exciting the coumarin at 409 nm now produces a blue fluorescence signal at 447 nm. This blue signal can be readily observed under a microscope and can also be detected as an increase in the blue channel readout on fluorescent
microplate readers.

The CCF2-AM and CCF4-AM substrates are essential assay components for the GeneBLAzer® platform. These substrates are fully compatible with flow cytometry, speeding the time to clone selection. Ratiometric analysis of the blue and green signals reduces well-to-well variation due to differences in cell numbers and substrate loading, leading to high Z´-factor values and low coefficients of variation (CVs).

CCF2-AM and CCF4-AM differ by two carbons in the bridge linking the coumarin moiety to the lactam ring. Both are in the membrane-permeable, esterified forms, and can be used for assays in intact cells. CCF4-AM has better solubility properties (soluble for >24 hours) than CCF2-AM and is thus best suited for screening applications. In addition, CCF4-AM has slightly better FRET and thus slightly lower background than CCF2-AM.

CCF2-FA is essentially the CCF2 substrate without the esters found in the AM version. CCF2-FA is de-esterified and used in cell lysate applications, bypassing loading across the cell membrane and de-esterification steps. Cell lysates are the preferred method for applications using cells that contain a cell wall. CCF2-FA can also be used as a control to acquire the excitation and emission spectra for CCF2-AM and CCF4-AM.

EnzChek™ Cellulase Substrate, blue fluorescent, 339/452 (Invitrogen™)

The EnzChek® cellulase substrate was developed for simple and rapid quantitation of cellulase. The assay can be completed in 30 minutes or less. Simply mix the enzyme with the EnxChek® cellulase substrate working solution, incubate, and read the results using excitation/emission maxima ~339/452 nm. In contrast to other more complex, multistep assays, the EnzChek® cellulase substrate is perfect for use in a high-throughput environment. This fluorescence substrate is highly sensitive, with a detection limit as low as 40 µU/mL cellulase. It is also possible to use this substrate in a colorimetric assay, albeit with reduced sensitivity.