In this product minireview we discuss the basic science of Luminol Chemiluminescence, its applications and the main areas of innovation in the field. Just Click the link below to download!
The general mechanism of luminol light emission:
In water and aqueous mixtures of alcohols or organic solvents a few conditions are necessary for light emission: a base, hydrogen peroxide and an oxidizing agent.
A base promoted deprotonation of luminol generates a negatively charged intermediate wich is in equilibrium with its corresponding “enolic” form. The latter reacts with molecular oxygen or hydrogen peroxide (depending on the conditions) to form an unstable endoperoxide, which decomposes releasing an aminophtalate dianion in an electronically excited state (and a molecule of nitrogen gas). The newly formed di-anion undergoes relaxation from the excited states (T1, S1) to the ground state (S0), thus emitting light.
(Biosynth Cat. No. L-8600)
Luminol-based Western blot assays use specific antibodies coupled to horseradish peroxidase (HRP) for immunodetection methods. HRP can catalyze the chemiluminescent oxidation of luminol in the presence of hydrogen peroxide. This transformation is used in ultra-sensitive ELISA assays or ECL detection.
Biosynth’s manufacturing process of Luminol ensures that highest purity and lot-to-lot consistency is achieved. Therefore it is the product of choice for manufacturers in and high-end immunological assays.
The review covers basic science of Luciferase enzymes, mechanistic aspects of FLuc bioluminescence, S.A.R of Luciferins and top-notch discoveries in the field. Particular attention is given to the highly promising area of in vivo bioluminescence. Just Click the link below to download!
Luciferin is a substrate for firefly luciferase used for imaging the expression of the luc marker gene and luciferase-fusion conjugates in living cells, tissues and organisms. Luciferin is extensively utilized in reporter gene assays, immunoassays and hygiene monitoring.
D-Luciferin Firefly L-8200
D-Luciferin Firefly, potassium salt, L-8220
D-Luciferin Firefly, sodium salt monohydrate, L-8240
L-Luciferin, potassium salt, L-8280
D-Luciferin 1-(4,5-dimethoxy-2-nitrophenyl)ethyl ester, L-8300
DL-Luciferin 1-(4,5-dimethoxy-2-nitrophenyl)ethyl ester, L-8305
Lucigenin is a diacridinium compound typically available in the form of a nitro salt. By reaction with superoxide anion it emits a blue-green light. Lucigenin-dependent luminescence can detect the formation of the important physiological superoxide radical which represents the main primary radical formed by the cell membranes and mitochondria. As with other chemiluminescent detection methods Lucigenin detection is extremely sensitive. In addition, Lucigenin can find applications for anion detection, in particular chloride which is able to quench its fluorescence and it has been employed for measuring chloride concentration in liposomes and artificial vesicles.
(CTZ) consists of an imidazopyrazinone nucleus and three lateral groups
connected to it. CTZ forms a stable solution in methanol, but is poorly soluble
and unstable in aqueous solutions, which makes its use in biological applications
more difficult. However, methods for preparation of aqueous solutions have been
developed, using NaOH and NH4OH or hydroxylpropyl-beta-cyclodextrin.
Coelenterazine, native C-7001 and C-7002
The luminescence of Coelenterazine: Light emission following decomposition of CTZ can be
observed through a bio-chemiluminescence. When O2 binds to the C-2 position of
the imidazopyrazinone core, a formation of peroxide is observed. The peroxide
produces a four-membered ring, dioxetanone, which breaks down to produce the
amide anion of coelenteramide and CO2. The anion is in a high energy state and
light emission is observed as the excited anion relaxes to a ground state.
Upon a chemiluminescence reaction of CTZ, a number of light emitters can be formed
from the coelenteramide anion, depending on the solvents used. In
bioluminescence, a luciferase catalyzes the reaction and the maximum wavelength
of the CTZ emission differs, depending on the luciferases utilized – for
example, the emission peak obtained with Oplophorus luciferase is 462 nm, while
with Renilla luciferase is 485 nm.
Biosynth offers a range of Coelenterazines:
Coelenterazine, native, CAS [55779-48-1], C-7001 and C-7002
Coelenterazine h, CAS [50909-86-9], C-7004
Coelenterazine 400a, CAS [70217-82-2], C-7011
Coelenterazine e, CAS [114496-02-5], C-7020
This minireview highlights the fascinating chemistry of dioxetanes and their exceptional light emission properties. Special attention is given to recent discoveries in the field which changed the game of chemiluminence. Just Click the link below to download!
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.
1. For example AquaSpark™ Peroxide Probe – one of the most sensitive H2O2 probes available.
2. For the detection of phosphatase activity we offer two product options: 1. AquaSpark™ Alkaline Phosphatase substrates that works best at alkaline pH values and is the ideal substrate for the commonly used (calf intestine) alkaline phosphatase. 2. AquaSpark™ Broad Range Phosphatase Substrate that works under a pH range from slightly acidic or neutral to alkaline – it is also a very good substrate for the AP from calf intestine but also a substrate for many other phosphatase isoenzymes.
3. As rapid and accurate identification of microorganisms remains an important challenge in microbiology we developed a range of AquaSpark™ enzyme substrates for rapid bacteria detection and identification.
Chemiluminescence and bioluminescence in nature
Chemiluminescence and bioluminescence (the analogous phenomenon in organisms) is observed when a molecule in a chemical reaction changes to an exited state and than emits the excitation energy as a photon. The Chemiluminescence is not an exotic phenomenon, actually a large number of organic materials emit small quantities of photons even at room temperature as a result of oxidation processes. Typically the wavelengths are of emitted light are between 400 - 700 nm, but can extend to the infrared region and UV.
The majority of chemiluminescence processes in nature result from redox reactions. An excitation energy of 160 - 320 kJ/mol is required to emit visible light by chemiluminescence. This energy needs to be available in a single reaction step, within a short time and within a relatively small reaction volume, so it is quenched by secondary reactions. (In reaction media fluorophores may be present that are capable to act as energy acceptors. Those molecules may absorb the excitation energy and show fluorescence light as a result).
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Please visit our web shop to see the full list of our chemiluminescence products: