The AquaSpark™ luminophor produces the strongest light you’ve ever experienced in any bio-assay. The luminescence is detectable with a luminometer (microplate reader with chemiluminescence capability) or on film. With the pioneering AquaSpark™ probes and substrates research tools become available that provide a previously unseen sensitivity.
AquaSpark™ - The new generation of chemiluminescence
that outshines all other detection methods.
AquaSpark™ molecules are the new generation of chemiluminescence probes for research and diagnostic applications. AquaSpark™ probes offer unique advantages as they can work as single agents, they have a higher efficiency and sensitivity over currently existing probes and they are especially designed to work under physiological conditions and can be used ex-vivo and in-vivo.
The first commercially available products based on the AquaSpark™ technology are AquaSpark™ Alkaline Phosphatase Substrate and AquaSpark™ Broad Range Phosphatase Substrate. They are the ideal substrates for chemiluminescene-based immunoassays (Western blot and ELISA) and they are suitable substrates for alkaline phosphatase activity blood serum tests (of ALP-related diseases). Both of them work in aqueous solutions, with the AquaSpark™ Broad Range Phosphatase Substrate even under neutral or slightly acidic pH conditions.
• They show very high light levels,
• have the lowest background,
• enable low detection limits,
• work as single reagent without enhancers,
• allow easy assay designs,
• enable cost-effective testing,
• are water-soluble,
• and work under physiological conditions.
AquaSpark ™ Alkaline Phosphatase Substrate (Cat. No. A-8164_P00)
• works at alkaline pH values
• is the ideal substrate for commonly used alkaline phosphatase (calf intestine)
AquaSpark ™ Broad Range Phosphatase Substrate (Cat. No. A-8163_P00)
• works under slightly acidic to alkaline pH values
• is a sensitive substrate for a wider range of phosphatase enzymes, including alkaline phosphatases.
beta-galactosidase (EC 220.127.116.11, shortly beta-Gal) catalyzes the hydrolysis of beta-d-galactoside in the presence of water to galactose and alcohol. beta-Gal has a molecular weight of 540,000 and is composed of four identical subunits of MW 135,000, each with an independent active site.The enzyme has divalent metals as cofactors, with chelated Mg2+ ions required to maintain active site conformation. The molecule contains numerous sulfhydryl groups and is glycosylated.
This type of enzyme is found widespread in many microorganisms, plants and animals. In E. coli for instance, the beta-Gal protein is encoded by the lacZ gene. The enzyme is often used as a reporter for monitoring gene expression. beta-Gal also has good characteristics when conjugated to antibody molecules or streptavidin for use in ELISA kits. In addition, the enzyme is used to determine lactose in biological fluids and it is employed in food processing operations. Traditionally, beta-Gal activity is tested using the X-Gal or ONPG chromogenic substrates. But with the pioneering AquaSpark™ beta-D-galactoside a substrate for beta-Gal detection becomes available that provides a previously unseen sensitivity.
Reference: Enzyme Modification and Conjugation. Greg T. Hermanson, in Bioconjugate Techniques (Third Edition), 2013
Image: NCBI.Madej T, Lanczycki CJ, Zhang D, Thiessen PA, Geer RC, Marchler-Bauer A, Bryant SH. " MMDB and VAST+: tracking structural similarities between macromolecular complexes.Nucleic Acids Res. 2014 Jan; 42(Database issue):D297-303
AquaSpark™ beta-D-galactoside, 10 mM in DMSO, Patent pending (Cat. No. A-8169_P00)
AquaSpark™ beta-D-galactoside is typically used at 10 – 50 µM final concentration. Biosynth provides AquaSpark™ beta-D-galactoside as a 10 mM stock solution in dimethyl sulfoxide (DMSO) for convenience and improved storage stability.
AquaSpark™ Peroxide Probe is the most sensitive probe for hydrogen peroxide (H2O2) detection available. The mechanism of chemiluminescence using AquaSpark™ relies on a boronic ester cleavable group which is oxidized in the presence of H2O2 in order to trigger the chemiluminescent reaction.
• Hydrogen peroxide detection. Excessive H2O2 production in cells and tissues may be the consequence of pathological conditions like inflammation, cancer, cardiovascular and neurodegenerative diseases. H2O2 detection in vitro and in vivo is of high importance in research and diagnostics.
• AquaSpark™ Peroxide Probe permits the detection of any H2O2 releasing oxidase activity, such as L-amino acid oxidase, monoamine oxidase, uiricase (urate oxidase), nucleoside oxidase and others.
•Glucose quantitation. Glucose oxidase (GOx) is a H2O2 producing oxidase and is widely used for the quantitation of free glucose. AquaSpark™ Peroxide Probe (A-8170_P00) coupled to GOx is ideal for the quantitation of glucose levels in body fluids (sera, plasma), foods, fermentation media, or vegetal raw material.
AquaSpark™ Peroxide Probe, 10 mM in DMSO, Patent pending (Cat. No. A-8170_P00)
• the most sensitive probe for H2O2 detection available
• an extremely versatile probe for oxidase detection applicable in a wide range of biocompatible buffers
AquaSpark™ alpha-D-glucoside (A-8171_P00) is the first single-compound chemiluminescent substrate commercially available for any assay or analytical method based on the detection of alpha-glucosidase activity.
Monitoring alpha-glucosidase is clinically relevant because is correlates with various diseases; for example lack of alpha- glucosidase leads to glycogen storage diseases (Pompe disease). Furthermore, numerous alpha-glucosidase inhibitors are used to treat type II diabetes, and also as antiviral agents.
In microbiology, alpha-glucosidase probes can be used for example for the detection of:
• Saccharomyces cerevisiae
• Cronobacter sakazakii
• Staphylococcus aureus
• Enterobacter spp.
• Bacillus stearothermophilus
• All species expressing alpha glucosidase
AquaSpark™ alpha-D-glucoside, 10 mM in DMSO, Patent pending (Cat. No. A-8171_P00)
• highly sensitive detection of pathogens for a wide range of applications
• the first commercially available chemiluminescent substrate for the detection of alpha-glucosidase
Esterases and lipases are widespread in cells and play a key role in energy metabolism and in the recycling of cellular components. In microbiology, C8-esterase activity can be used to distinguish between Salmonella strains and other enteric bacteria. Likewise, Pseudomonas species can be also detected, since these organisms produce extracellular lipases. Upon hydrolytic cleavage by C8-esterase (or lipase) AquaSpark™ caprylate (A-8167_P00) spontaneously emits a strong green light that allows the sensitive detection of C8-esterase and lipase activity in vitro and in vivo.
AquaSpark™ caprylate, 10 mM in DMSO, Patent pending (Cat. No. A-8167_P00)
• sensitive detection of C8-esterase and lipase activity in vitro and in vivo
AquaSpark™ beta-D-glucuronide (A-8175_P00) is the first single-compound luminogenic substrate commercially available for highly sensitive detection of Escherichia coli, GUS reporter gene assays and further analytical methods based on the detection of beta-glucuronidase. Upon hydrolytic cleavage AquaSpark™ beta-D-glucuronide spontaneously emits a green light that allows for the sensitive detection of beta-glucuronidase activity in almost every biological sample.
Escherichia coli is the most important indicator of fecal contamination of water, beverages and food. Living cells of E. coli can be reliably detected with media that contain a substrate for beta-glucuronidase, an enzyme that occurs in the microbial world mainly but with a few exceptions in E. coli. AquaSpark™ beta-D-glucuronide is a revolutionary luminescent substrate for the rapid E. coli identification.
The most popular reporter gene assays in plant molecular biology are also based on the expression of the beta-glucuronidase (GUS). GUS activity may be detected with chemiluminescence by using the same AquaSpark™ beta-D-glucuronide probe (A-8175_P00) instead of the less sensitive fluorescence or chromogenic methods.
AquaSpark™ beta-D-glucuronide, 10 mM in DMSO, Patent pending (Cat. No. A-8175_P00)
• bright chemiluminescence in aqueous solutions
• E. coli identification after only 6 hours cultivation
• easy handling
• more sensitive than MUG or chromogenic substrates
For the detection of nitroreductase activity and co-factor NADH in a wide range of different species.
The AquaSpark™ 510 Nitroreductase/NADH Probe (CL-NTR) is a new probe for the selective chemiluminescent detection of nitroreductase (NTR) activity.
It can be used for the detection of endogenous NTR activity in bacteria or animal models, e.g. in hypoxia tumor models. The same probe can also be used to determine NADH concentration in vitro when used together with a defined NTR enzyme concentration.
The NTR porbe is characterized by a high selectivity towards Nitroreductase and cofactor NADH. Other reducing agents do not interfere with the detection system.
A. Detection of bacterial nitroreductase: The lysis of NTR-positive bacteria with reagents such as polymyxin and lysozyme releases the NTR enzyme activity. The NTR activity can be subsequently
determined very precisely with the AquaSpark™ probe.
B. Quantification of NADH - chemiluminescence kineticprofile:The chemiluminescence level of AquaSpark™ Nitroreductase/NADH probe (CL-NTR) is very high immediately after addition of nitroreductase and reaches its relative maximum point after about 2 min. The luminescence level remains high for at least 6 to 10 min, depending on NADH and NTR concentrations. During this time chemiluminescent measurements deliver a superior signal-to-background ratio and consequently NADH measurements of highest sensitivity and accuracy.
AquaSpark™ 510 Nitroreductase/NADH Probe, lyophilized, Patent pending (Cat. No. A-8175_P00)
• high sensitivity, due to high signal-to-background ratio
• high selectivity
• easy handling
• rapid results
Multiple challenges for microbial diagonsitcs such as the growing number of foodborne illnesses and emerging antibiotic resistances of bacteria require breakthrough ideas and innovations in more rapid pathogen detection solutions to be brought to the market.
Co-founded by Biosynth AG, Ramot at Tel Aviv University and its management, NEMIS Technologies has acquired the worldwide and exclusive license for the use of the AquaSpark™ technology in microbiological applications for the development of rapid, precise, easy-to-use and low-cost diagnostic solutions.
For further information on AquaSpark™ in this field please contact NEMIS: BD@nemistech.com