A.G. Scientific, Inc. is excited to be able to offer our clients a large selection of fluorescent dyes, reagents and imaging kits useful in fluorescent microscopy, in situ hybridization, flow cytometry, multi-well assays, content screening, reporter assays, and many more.

THE ROLE OF LIGHT

Light plays a primary role in the universe. On Earth, primary producers use the photosynthetic pathway to convert light energy into chemical energy, which can then be used as food. Many other organisms are able to produce their own light as a method of communication, to see where there's no light, aid predation, mating, or as a defensive weapon.

In the case of organisms living in the deep sea, they have developed some rather sophisticated ways to produce light through a marvel known as bioluminescence. The phenomenon is made possible by a chemical reaction between two biochemicals, Luciferase (the enzyme) and Luciferin (the substrate), as well as the required oxygen and ATP energy input.

Biologists and engineers are studying the circumstances for the activation of luciferase, the enzyme involved in bioluminescence, to better understand how human physiology functions and how diseases develop. We can thank Aristotle for discovering it. Robert Boyle, a British scientist, conducted what would become the first modern scientific study by analyzing the properties of luminescence. French biologist, Raphael Dubois, later thought of the reaction sequence between luciferins that emit light.

VISUALIZING CELLS IN THE LABORATORY

A chemist has a choice between assays, one usually depending on their preference for which biomarker is being analyzed or the amount of sensitivity required.

The improvements in assay technology over the past decade, most notably with microwell plates, are enabling faster, smaller, and more efficient detection measurements. Certainly one of the most significant additions to a biochemist's tool belt is the conveniently automated high-throughput screening (HTS) for a large number of compounds. In the laboratory, Luciferase is most commonly used biochemical in the HTS Assays.

The goal of every HTS Assay is to investigate intracellular events by monitoring gene transcription. This may include observing internal controls through the simple and efficient dual-luciferase assay. Most commonly, this technique is used for GPCR and nuclear receptor assays.

To obtain best results, your enzyme of choice (i.e. Luciferase) must be consistent between assays. It is therefore best to aim for consistency within your research, as well as to the reproducibility of your work, to use stable forms of your labeling reagent. The main advantage of using Luciferase purchased from A.G. Scientific is that it is 100X more sensitive and is significantly faster than most of the dye assays used to look at cellular metabolism.

The main advantage of using Luciferase purchased from A.G. Scientific is that it is 100X more sensitive and significantly faster than most of the dye assays used to look at cellular metabolism1.


Laser Capture

hts_assay_example

Image Research: Trends in Biotechnology. (2004). [1]

Tubulin labeled with fluorescein, for example, can be integrated into the microtubules of spindles seen best during mitosis. When a laser hits a certain region of the spindle, the labeled tubulin is illuminated so that its movements can be tracked and studied. In principle, the same technique can be applied to any protein.

Overall, the most widely used fluorescent protein overall among biologists is the green fluorescent protein, derived from the jellyfish Aequoria victoria. One of its many uses is as a reporter molecule capable of monitoring gene expression. We will have to report more on that later as news breaks.

If you are currently seeking a primary supplier for your research, our experienced staff can help assist you in placing your bulk order.

Luciferase enzymes can be used in:

  1. Cytotoxic screens which measure ATP concentrations and cell viability
  2. Kinase Activity screens since all kinases consume ATP
  3. Real-time detection of ATPase activity

A.G. Scientific Assays, Reagents, and Kits

SOURCES/REFERENCES

[1] Roda, Aldo, Patrizia Pasini, Mara Mirasoli, Elisa Michelini, and Massimo Guardigli. "Biotechnological Applications of Bioluminescence Adn Chemiluminescence." TRENDS in Biotechnology 22.6 (2004): 295-303.

[2] Seminario-Vidal, Lucia, Eduardo R. Lazarowski, and Seiko F. Okada. "Assessment of Extracellular ATP."Bioluminescence, Methods in Molecular Biology 574 (2009): 25-36.

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  1. CRT Inhibitor, iCRT5 chemical structure | CAS 18623-44-4  | AG Scientific, Inc.
    iCRT5 (CRT Inhibitor)
    C-2273

    Starting at $91.98

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