By Sung Bae Kim
This quantity offers precise laboratory protocols in regards to the 3 significant path applied sciences luciferases, effective optical probes, and purposes to visualizing molecular occasions in dwelling subjects. Bioluminescence: equipment and Protocols, 3rd Edition guides readers via chapters on elements of bioluminescent probes, and fabrication of bioluminescent probes, purposes to residing topics and instrumentations. Written within the hugely winning Methods in Molecular Biology series layout, chapters contain introductions to their respective themes, lists of the mandatory fabrics and reagents, step by step, without difficulty reproducible laboratory protocols, and tips about troubleshooting and warding off identified pitfalls.
Authoritative and useful, Bioluminescence: tools and Protocols, 3rd Edition is an invaluable supplement to the 1st and moment version for brand new and skilled researchers alike.
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Extra resources for Bioluminescence: Methods and Protocols
2025 [M-H]−. 14 Synthesis of (E)-8-Benzyl-2-(4hydroxybenzyl)-6-(4hydroxystyryl) imidazo[1,2-a]pyrazin3(7H)-one (See Fig. 1 Compound ( 14)) (6-pi-OH-CTZ) Yield 23 % (yellow solid compound). 1 % formic acid. 09 (s, 2H). 8. 1661 [M-H]−. 1. Add a CH3OH solution of the respective CTZ derivative (1 mM, 200 μL) to a quartz cell and set in a SREX Fluorolog-3 fluorescence spectrophotometer (Model FL-3-11, Horiba Jobin Yvon, Kyoto, Japan). 2. Measure the chemiluminescence spectra at a scan rate of 1200 nm/min after injecting 2 mL of DMSO (see Note 11).
55 °C for 10 s for the annealing. 68 °C for 30 s for the extension. 1 cycle for the last extention step. 68 °C for 30 s. 3. 6 % TAE-agarose gel with suitable DNA size marker to analyze PCR products. 4. If a single discrete band around 100–300 bp is observed, cut the band from the gel and gel-purify the product using the gel purification kit (see Note 5). Molecular Cloning of Secreted Luciferases from Copepods 39 5. 1-TOPO vector (Life Technologies), then incubate for 5–15 min at room temperature, and transform into the competent cells, such as TOP10 or DH5α (Life Technologies).
10. Normalize the spectra in relative percentages to the maximal intensity (λmax) (see Fig. 3 and Note 11). 3 Application to Bioluminescent Capsules ALuc® can be linked to a membrane localization signal (MLS). Any protein including a fluorescent protein can be inserted between ALuc and MLS. This type of probes are named “bioluminescent capsule,” the fabrication procedure of which is briefly described in the caption of Fig. 2. The basic concept was described in our previous study  (see Fig. 2).